Oligoribonucleotide inhibitors of Nrf2 and methods of use thereof for treatment of cancer

ABSTRACT

The invention provides novel double stranded oligoribonucleotides that inhibit the Nrf2 gene. The invention also provides a pharmaceutical composition comprising one or more such oligoribonucleotides, and a vector capable of expressing the oligoribonucleotide. The present invention also relates to methods and compositions for treating or preventing the incidence or severity of a cancerous disease, particularly various lung cancers.

This application is a divisional of U.S. Ser. No. 12/077,414 filed Mar. 19, 2008, now U.S. Pat. No. 7,812,002, issued Oct. 12, 2010, which claims the benefit of U.S. Provisional Application No. 60/937,135, filed Jun. 26, 2007 and of U.S. Provisional Application No. 60/919,413, filed Mar. 22, 2007, which contents of all of which are hereby incorporated by reference into this application.

Throughout this application various patent and scientific publications are cited. The disclosures for these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains.

This application contains nucleotide and/or amino acid sequences which are present in the file named “20120603_(—)77688_AZ_SubstituteSequenceListing.txt,” which is 348 KB in size, and was created on Jun. 3, 2012 in the IBM-PCT machine format and has an operating system compatibility with MS-Windows, which is contained in the text file filed Jun. 13, 2012 as part of this application.

BACKGROUND OF THE INVENTION siRNAs and RNA Interference

RNA interference (RNAi) is a phenomenon involving double-stranded (ds) RNA-dependent gene specific posttranscriptional silencing. Originally, attempts to study this phenomenon and to manipulate mammalian cells experimentally were frustrated by an active, non-specific antiviral defense mechanism which was activated in response to long dsRNA molecules; see Gil et al. 2000, Apoptosis, 5:107-114. Later it was discovered that synthetic duplexes of 21 nucleotide RNAs could mediate gene specific RNAi in mammalian cells, without the stimulation of the generic antiviral defense mechanisms (see Elbashir et al. Nature 2001, 411:494-498 and Caplen et al. Proc Natl Acad Sci 2001, 98:9742-9747). As a result, small interfering RNAs (siRNAs), which are short double-stranded RNAs, have become powerful tools in attempting to understand gene function.

Thus, RNA interference (RNAi) refers to the process of sequence-specific post-transcriptional gene silencing in mammals mediated by small interfering RNAs (siRNAs) (Fire et al, 1998, Nature 391, 806) or microRNAs (miRNAs) (Ambros V. Nature 431:7006, 350-355 (2004); and Bartel D P. Cell. 2004 Jan. 23; 116(2): 281-97 MicroRNAs: genomics, biogenesis, mechanism, and function). The corresponding process in plants is commonly referred to as specific post-transcriptional gene silencing or RNA silencing and is also referred to as quelling in fungi. An siRNA is a double-stranded RNA molecule which down-regulates or silences (prevents) the expression of a gene/mRNA of its endogenous (cellular) counterpart. RNA interference is based on the ability of dsRNA species to enter a specific protein complex, where it is then targeted to the complementary cellular RNA and specifically degrades it. Thus, the RNA interference response features an endonuclease complex containing an siRNA, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having a sequence complementary to the antisense strand of the siRNA duplex. Cleavage of the target RNA may take place in the middle of the region complementary to the antisense strand of the siRNA duplex (Elbashir et al 2001, Genes Dev., 15, 188). In more detail, longer dsRNAs are digested into short (17-29 bp) dsRNA fragments (also referred to as short inhibitory RNAs—“siRNAs”) by type III RNAses (DICER, DROSHA, etc., Bernstein et al., Nature, 2001, v. 409, p. 363-6; Lee et al., Nature, 2003, 425, p. 415-9). The RISC protein complex recognizes these fragments and complementary mRNA. The whole process is culminated by endonuclease cleavage of target mRNA (McManus & Sharp, Nature Rev Genet, 2002, v. 3, p. 737-47; Paddison & Hannon, Curr Opin Mol. Ther. 2003 June; 5(3): 217-24). For information on these terms and proposed mechanisms, see Bernstein E., Denli A M. Hannon G J: 2001 The rest is silence. RNA. I; 7(11): 1509-21; Nishikura K.: 2001 A short primer on RNAi: RNA-directed RNA polymerase acts as a key catalyst. Cell. 116; 107(4): 415-8 and PCT publication WO 01/36646 (Glover et al).

The selection and synthesis of siRNA corresponding to known genes has been widely reported; see for example Chalk A M, Wahlestedt C, Sonnhammner E L. 2004 Improved and automated prediction of effective siRNA Biochem. Biophys. Res. Commun. June 18; 319(1): 264-74; Sioud M, Leirdal M., 2004, Potential design rules and enzymatic synthesis of siRNAs, Methods Mol. Biol.; 252:457-69; Levenkova N, Gu Q, Rux J J. 2004, Gene specific siRNA selector Bioinformatics. 112; 20(3): 430-2. and Ui-Tei K, Naito Y, Takahashi F, Haraguchi T, Ohki-Hamazaki H, Juni A, Ueda R, Saigo K., Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference Nucleic Acids Res. 2004 I 9; 32(3):936-48. See also Liu Y, Braasch D A, Nulf C J, Corey D R. Efficient and isoform-selective inhibition of cellular gene expression by peptide nucleic acids, Biochemistry, 2004 I 24; 43(7):1921-7. See also PCT publications WO 2004/015107 (Atugen) and WO 02/44321 (Tuschl et al), and also Chiu Y L, Rana T M. siRNA function in RNAi: a chemical modification analysis, RNA 2003 September; 9(9):1034-48 and I U.S. Pat. Nos. 5,898,031 and 6107094 (Crooke) for production of modified/more stable siRNAs.

Several groups have described the development of DNA-based vectors capable of generating siRNA within cells. The method generally involves transcription of short hairpin RNAs that are efficiently processed to form siRNAs within cells. Paddison et al. PNAS 2002, 99:1443-1448; Paddison et al. Genes & Dev 2002, 16:948-958; Sui et al. PNAS 2002, 8:5515-5520; and Brummelkamp et al. Science 2002, 296:550-553. These reports describe methods to generate siRNAs capable of specifically targeting numerous endogenously and exogenously expressed genes.

Several studies have revealed that siRNA therapeutics are effective in viva in both mammals and in humans. Bitko et al., have shown that specific siRNA molecules directed against the respiratory syncytial virus (RSV) nucleocapsid N gene are effective in treating mice when administered intranasally (Bitko et al., Nat. Med. 2005, 11(1):50-55). For reviews of therapeutic applications of siRNAs see for example, Batik (Mol. Med. 2005, 83: 764-773); Chakraborty (Current Drug Targets 2007 8(3):469-82) and Dykxhoom, et al (Gene Therapy 2006, 13, 541-552). Furthermore, a phase I clinical study with short siRNA molecule that targets the VEGFR1 receptor for the treatment of Age-Related Macular Degeneration (AMD) has been conducted in human patients. In studies such siRNA administered by intravitreal (intraocular) injection was found effective and safe in 14 patients tested (Kaiser, Am J Ophthalmol. 2006 142(4):660-8).

The Nrf2 Gene and Polypeptide

(gi|166295208|ref|NM_(—)006164.3| Homo sapiens Nuclear Factor (Erythroid-Derived 2)-like 2 (NFE2L2):

Nuclear factor erythroid-2 related factor 2 (Nrf2), a cap-and-collar basic leucine zipper transcription factor, positively regulates a transcriptional program that maintains cellular redox homeostasis and protects cells from oxidative insult, including insult from chemotherapeutic agents (Rangasamy T, et al. J Clin Invest 114, 1248 (2004)). Nrf2 activates transcription of its target genes through binding specifically to the antioxidant-response element (ARE) found in those gene promoters. The Nrf2-regulated transcriptional program includes a broad spectrum of genes, including antioxidants such as heme oxygenase-1, superoxide dismutase, glutathione reductase (GSR), glutathione peroxidase, thioredoxin, thioredoxin reductase, peroxiredoxins (PRDX).

Lung Cancer:

Lung cancer is a cancer that forms in tissues of the lung, usually in the cells lining air passages. The two main types are small cell lung cancer and non-small cell lung cancer. These types are diagnosed based on the morphology of the cells under a microscope. It is the most lethal of all cancers worldwide, responsible for up to 3 million deaths annually. In non-small cell lung cancer (NSCLC), results of standard treatment are poor except for the most localized cancers. Surgery is the most potentially curative therapeutic option for this disease; radiation therapy can produce a cure in a small number of patients and can provide palliation in most patients. Adjuvant chemotherapy may provide an additional benefit to patients with resected NSCLC. In advanced-stage disease, chemotherapy offers modest improvements in median survival, though overall survival is poor. Chemotherapy has produced short-term improvement in disease-related symptoms.

WO 2006/128041 discloses specific siRNA molecules for Nrf2 and its use for treating any cancer, preferably lung and kidney cancers. US20020164576 discloses a method of inhibiting tumor growth (preferably a lymphoma cancer) using antisense molecules directed to Nrf2 or specific antibodies. US20070042418 discloses the use of siRNA molecules for Nrf2 for treating cancer.

Despite the evident progress, there remains a continued need for improved molecules in particular improved siRNA compounds able to treat cancerous diseases, particularly lung cancers.

SUMMARY OF THE INVENTION

The invention provides novel double stranded oligoribonucleotides that inhibit the Nrf2 gene. The invention also provides a pharmaceutical composition comprising one or more such oligoribonucleotides, and a vector capable of expressing the oligoribonucleotide. The present invention also relates to methods and compositions for treating or preventing the incidence or severity of a cancerous disease, particularly various lung cancers. The methods and compositions involve administering to a mammal in need of such treatment a prophylactically or therapeutically effective amount of one or more compounds which down-regulate expression of the Nrf2 gene, particularly novel small interfering RNAs (siRNAs), small molecule inhibitors of Nrf2 as described herein or antibodies to Nrf2 polypeptide.

In certain embodiments the present invention provides a compound having the structure:

5′ (N)_(x)-Z 3′ (antisense strand) 3′ Z′-(N′)_(y) 5′ (sense strand) wherein each of N and N′ is a ribonucleotide which may be independently modified or unmodified in its sugar residue; wherein each of (N)_(x) and (N′)_(y) is an oligomer in which each consecutive N or N′ is joined to the next N or N′ by a covalent bond; wherein each of x and y is an integer between 19 and 40; wherein each of Z and Z′ may be present or absent, but if present is 1-5 consecutive nucleotides covalently attached at the 3′ terminus of the strand in which it is present; and wherein the sequence of (N)_(x) is set forth as any one of SEQ ID NOS 504 to 1006 or 1507 to 2006, and the sequence of (N′)_(y) is set forth as the complementary sequence. The sequence of (N′)_(y) is a complementary sequence set forth as SEQ ID NOS: 1 to 503 or 1007 to 1506, respectively.

In some embodiments the covalent bond joining each consecutive Nor N′ is a phosphodiester bond. In various embodiments all the covalent bonds are phosphodiester bonds.

In various embodiments the compound comprises ribonucleotides wherein x=y and each of x and y is 19, 20, 21, 22 or 23. In some embodiments x=y=23. In other embodiments x=y=19.

In some embodiments the compound is blunt ended, for example wherein both Z and Z′ are absent. In an alternative embodiment, the compound comprises at least one 3′ overhang, wherein at least one of Z or Z′ is present. Z and Z′ can independently comprise one or more covalently linked modified or non-modified nucleotides, for example inverted dT or dA; dT, LNA, mirror nucleotide and the like. In some embodiments each of Z and V are independently selected from dT and dTdT.

In some embodiments Nor N′ comprises a modification in the sugar residue of one or more ribonucleotides. In other embodiments the compound comprises at least one ribonucleotide modified in the sugar residue. In some embodiments the compound comprises a modification at the 2′ position of the sugar residue. In some embodiments the modification in the 2′ position comprises the presence of an amino, a fluoro, an alkoxy or an alkyl moiety. In certain embodiments the 2′ modification comprises a methoxy moiety. A presently preferred modification is a 2′ methoxy of the sugar residue (2′-O-methyl; 2-O-Me; 2′-O—CH₃).

In some embodiments the compound comprises modified alternating ribonucleotides in one or both of the antisense and the sense strands. In certain embodiments the compound comprises modified alternating ribonucleotides in the antisense and the sense strands. In other embodiments the compound comprises modified alternating ribonucleotides in the antisense strand only. In certain embodiments the middle ribonucleotide of the antisense strand is not modified; e.g. ribonucleotide in position 10 in a 19-mer strand or position 12 in a 23-mer strand.

In additional embodiments the compound comprises modified ribonucleotides in alternating positions wherein each N at the 5′ and 3′ termini of (N)_(x) are modified in their sugar residues, and each N′ at the 5′ and 3′ termini of (N′)_(y) are unmodified in their sugar residues. In some embodiments, neither (N)_(x) nor (N′)_(y) are phosphorylated at the 3′ and 5′ termini. In other embodiments either or both (N)_(x) and (N′)_(y) are phosphorylated at the 3′ termini. Preferably the siRNA molecule is either phosphorylated at the 3′ termini of both sense and anti-sense strands, or non-phosphorylated at all; similar results are obtained for both siRNAs. The siRNA molecules used in the in vitro experiments were phosphorylated at the 3′ termini of both sense and anti-sense strands while the siRNA molecules used in the in vivo experiments were non-phosphorylated at all.

More specifically, the present invention provides methods and compositions for treating a patient suffering from a cancerous disease, (e.g. lung cancer, breast cancer, cervical cancer, colon cancer, gastric cancer, kidney cancer, leukemia, liver cancer, lymphoma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, sarcoma, skin cancer, testicular cancer, and uterine cancer). In a particular embodiment, the cancer is lung cancer such as non-small-cell lung carcinoma (NSCLC) or small-cell lung carcinoma. The methods of the invention comprising administering to the patient one or more compounds which down-regulate expression of the Nrf2 gene, particularly siRNAs that inhibit Nrf2, typically as a pharmaceutical composition, in a therapeutically effective dose so as to thereby treat the patient.

Still further, the invention relates to a method for treating or preventing the incidence or severity of a cancerous disease, particularly a lung disease in a patient comprising administering to the patient a composition comprising an effective amount of naked siRNA molecules. Preferably, the naked siRNA molecules are applied directly to lung for example via aerosol delivery which has the potential for delivering high concentrations of the therapeutic molecule to the inner lung.

Still further, the invention relates to the use of a therapeutically effective dose of an oligonucleotide for the preparation of a composition for treating a subject suffering from a cancerous disease, preferably lung cancer, wherein the oligonucleotide is set forth as any one of SEQ ID NOS:1-2006.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1: A549-C8 luciferase cells were transfected with non-targeting luciferase siRNA or Nrf2 siRNA-1 (sense strand SEQ ID NO: 10) at 20 nM concentration. Seventy two hours post transfection, cells were harvested and total RNA was isolated. Nrf2 knockdown as well as expression of two Nrf2 dependent target genes (GCLM and HO-1) were analyzed by real time RT-PCR.

FIG. 2: SCID-Beige mice injected i.v. with ARE-luciferase reporter tumor cells were inhaled with Nrf2 siRNA (sense strand SEQ ID NO: 10) twice during the 4th week of lung tumor growth. Control mice were inhaled with GFP siRNA. Mice were imaged before and after siRNA inhalation.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates generally to compounds which down-regulate expression of the Nrf2 gene particularly to novel small interfering RNAs (siRNAs), and to the use of these novel siRNAs in the treatment of various diseases and medical conditions in particular various forms of cancerous diseases. Such preferred siRNA compounds comprise sequences set forth as any one of SEQ ID NOS: 1 to 2006.

The inventors of the present invention have provided novel siRNAs to Nrf2 in order to treat any of the above diseases or disorders. Methods, molecules and compositions which inhibit Nrf2 are discussed herein at length, and any of said molecules and/or compositions may be beneficially employed in the treatment of a patient suffering from any of said conditions.

The present invention provides methods and compositions for inhibiting expression of a target Nrf2 gene in vivo. In general, the method includes administering oligoribonucleotides, such as small interfering RNAs (i.e., siRNAs) that are targeted to a particular Nrf2 mRNA and hybridize to, or interact with, the mRNAs under biological conditions (within the cell), or a nucleic acid material that can produce siRNA in a cell, in an amount sufficient to down-regulate expression of a target gene by an RNA interference mechanism. In particular, the subject method can be used to inhibit expression of the Nrf2 gene for treatment of a disease.

In accordance with the present invention, the siRNA molecules or inhibitors of the Nrf2 gene may be used as drugs to treat various pathologies accompanied by an elevated level of Nrf2 polypeptide.

The present invention provides double-stranded oligoribonucleotides (siRNAs), which down-regulate the expression of the Nrf2 gene. An siRNA of the invention is a duplex oligoribonucleotide in which the sense strand is derived from the mRNA sequence of the Nrf2 gene, and the antisense strand is complementary to the sense strand. In general, some deviation from the target mRNA sequence is tolerated without compromising the siRNA activity (see e.g. Czauderna et al 2003 Nucleic Acids Research 31(11), 2705-2716). An siRNA of the invention inhibits gene expression on a post-transcriptional level with or without destroying the mRNA. Without being bound by theory, siRNA may target the mRNA for specific cleavage and degradation and/or may inhibit translation from the targeted message.

As used herein, the term “Nrf2”, or “Nrf2 polypeptide” (gi|166295208|ref|NM_(—)006164.3| Homo sapiens nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) is defined as any homolog of the Nrf2 polypeptide having preferably 90% homology, more preferably 95% homology, and even more preferably 98% homology to the Nrf2 gene, as either full-length or a fragment or a domain thereof, as a mutant or the polypeptide encoded by a spliced variant nucleic acid sequence, as a chimera with other polypeptides, provided that any of the above has the same or substantially the same biological function as the Nrf2 polypeptide.

Generally, the siRNAs used in the present invention comprise a ribonucleic acid comprising a double stranded structure, whereby the double-stranded structure comprises a first strand and a second strand, whereby the first strand comprises a first stretch of contiguous nucleotides and whereby said first stretch is at least partially complementary to a target nucleic acid, and the second strand comprises a second stretch of contiguous nucleotides and whereby said second stretch is at least partially identical to a target nucleic acid, whereby said first strand and/or said second strand comprises a plurality of groups of modified nucleotides having a modification at the 2′-position whereby within the strand each group of modified nucleotides is flanked on one or both sides by a flanking group of nucleotides whereby the flanking nucleotides forming the flanking group of nucleotides is either an unmodified nucleotide or a nucleotide having a modification different from the modification of the modified nucleotides. Further, said first strand and/or said second strand may comprise said plurality of modified nucleotides and may comprises said plurality of groups of modified nucleotides.

The group of modified nucleotides and/or the group of flanking nucleotides may comprise a number of nucleotides whereby the number is selected from the group comprising one nucleotide to 10 nucleotides. In connection with any ranges specified herein it is to be understood that each range discloses any individual integer between the respective figures used to define the range including said two figures defining said range. In the present case the group thus comprises one nucleotide, two nucleotides, three nucleotides, four nucleotides, five nucleotides, six nucleotides, seven nucleotides, eight nucleotides, nine nucleotides and ten nucleotides.

The pattern of modified nucleotides of said first strand may be shifted by one or more nucleotides relative to the pattern of modified nucleotides of the second strand.

The modifications discussed above may be selected from the group comprising amino, fluoro, methoxy alkoxy, alkyl, amino, fluoro, chloro, bromo, CN, CF, imidazole, carboxylate, thioate, C₁ to C₁₀ lower alkyl, substituted lower alkyl, alkaryl or aralkyl, OCF₃, OCN, O—, S—, or N-alkyl; O-, S-, or N-alkenyl; SOCH₃; SO₂CH₃; ONO₂; NO₂, N₃; heterozycloalkyl; heterozycloalkaryl; aminoalkylamino; polyalkylamino or substituted silyl, as, among others, described in European patents EP 0 586 520 B1 or EP 0 618 925 B1.

The double stranded structure of the siRNA may be blunt ended, on one or both sides. More specifically, the double stranded structure may be blunt ended on the double stranded structure's side which is defined by the 5′-end of the first strand and the 3′-end of the second strand, or the double stranded structure may be blunt ended on the double stranded structure's side which is defined by at the 3′-end of the first strand and the 5′-end of the second strand.

Additionally, at least one of the two strands may have an overhang of at least one nucleotide at the 5′-end; the overhang may consist of at least one deoxyribonucleotide. At least one of the strands may also optionally have an overhang of at least one nucleotide at the 3′-end.

The length of the double-stranded structure of the siRNA is typically from about 17 to 21 and more preferably 18 or 19 bases. Further, the length of said first strand and/or the length of said second strand may independently from each other be selected from the group comprising the ranges of from about 15 to about 23 bases, 17 to 21 bases and 18 or 19 bases.

Additionally, the complementarily between said first strand and the target nucleic acid may be perfect, or the duplex formed between the first strand and the target nucleic acid may comprise at least 15 nucleotides wherein there is one mismatch or two mismatches between said first strand and the target nucleic acid forming said double-stranded structure.

Substantially complementary refers to complementarity of greater than about 84%, to another sequence. For example in a duplex region consisting of 19 base pairs one mismatch results in 94.7% complementarity, two mismatches results in about 89.5% complementarity and 3 mismatches results in about 84.2% complementarity, rendering the duplex region substantially complementary. Accordingly substantially identical refers to identity of greater than about 84%, compared to another sequence.

In some cases both the first strand and the second strand each comprise at least one group of modified nucleotides and at least one flanking group of nucleotides, whereby each group of modified nucleotides comprises at least one nucleotide and whereby each flanking group of nucleotides comprising at least one nucleotide with each group of modified nucleotides of the first strand being aligned with a flanking group of nucleotides on the second strand, whereby the most terminal 5′ nucleotide of the first strand is a nucleotide of the group of modified nucleotides, and the most terminal 3′ nucleotide of the second strand is a nucleotide of the flanking group of nucleotides. Each group of modified nucleotides may consist of a single nucleotide and/or each flanking group of nucleotides may consist of a single nucleotide.

Additionally, it is possible that on the first strand the nucleotide forming the flanking group of nucleotides is an unmodified nucleotide which is arranged in a 3′ direction relative to the nucleotide forming the group of modified nucleotides, and on the second strand the nucleotide forming the group of modified nucleotides is a modified nucleotide which is arranged in 5′ direction relative to the nucleotide forming the flanking group of nucleotides.

Further the first strand of the siRNA may comprise eight to twelve, preferably nine to eleven, groups of modified nucleotides, and the second strand may comprise seven to eleven, preferably eight to ten, groups of modified nucleotides.

The first strand and the second strand may be linked by a loop structure, which may be comprised of a non-nucleic acid polymer such as, inter alia, polyethylene glycol. Alternatively, the loop structure may be comprised of a nucleic acid.

Further, the 5′-terminus of the first strand of the siRNA may be linked to the 3′-terminus of the second strand, or the 3′-end of the first strand may be linked to the 5′-terminus of the second strand, said linkage being via a nucleic acid linker typically having a length between 10-2000 nucleobases.

In particular, the invention provides a compound having structure A:

5′ (N)_(x)-Z 3′ (antisense strand) 3′ Z′-(N′)_(y) 5′ (sense strand)

-   -   wherein each N and N′ is a ribonucleotide which may         independently be modified or unmodified in its sugar residue and         each of (N)_(x) and (N′)_(y) is oligomer in which each         consecutive N or N′ is joined to the next N or N′ by a covalent         bond;     -   wherein each of x and y is an integer between 19 and 40;     -   wherein each of Z and Z′ may be present or absent, but if         present is dTdT and is covalently attached at the 3′ terminus of         the strand in which it is present;     -   and wherein the sequence of (N)_(x) is set forth as any one of         SEQ ID NOS 504 to 1006 or 1507 to 2006, and the sequence of         (N′)_(y) is set forth as the complementary sequence. The         sequence of (N′)_(y) is a complementary sequence set forth as         SEQ ID NOS: 1 to 503 or 1007 to 1506, respectively.

It will be readily understood by those skilled in the art that the compounds of the present invention consist of a plurality of nucleotides, which are linked through covalent linkages. Each such covalent linkage may be a phosphodiester linkage, a phosphothioate linkage, or a combination of both, along the length of the nucleotide sequence of the individual strand.

Other possible backbone modifications are described inter alia in U.S. Pat. Nos. 5,587,361; 6,242,589; 6,277,967; 6,326,358; 5,399,676; 5,489,677; and 5,596,086.

In particular embodiments, x and y are preferably an integer between about 19 to about 27, most preferably from about 19 to about 23. In a particular embodiment of the compound of the invention, x may be equal to y (viz., x=y) and in preferred embodiments x=y=19 or x=y=21. In a particularly preferred embodiment x=y=19.

In one embodiment of the compound of the invention, Z and Z′ are both absent; in another embodiment one of Z or Z′ is present.

In one embodiment of the compound of the invention, all of the ribonucleotides of the compound are unmodified in their sugar residues.

In preferred embodiments of the compound of the invention, at least one ribonucleotide is modified in its sugar residue, preferably a modification at the 2′ position. The modification at the 2′ position results in the presence of a moiety which is preferably selected from the group comprising amino, fluoro, methoxy, alkoxy and alkyl groups. In a presently most preferred embodiment the moiety at the 2′ position is methoxy (2′-0-methyl).

In preferred embodiments of the invention, alternating ribonucleotides are modified in both the antisense and the sense strands of the compound. In particular the siRNA used in the Examples has been such modified such that a 2′ O-Me group was present on the first, third, fifth, seventh, ninth, eleventh, thirteenth, fifteenth, seventeenth and nineteenth nucleotide of the antisense strand, whereby the very same modification, i.e. a 2′-O-Me group was present at the second, fourth, sixth, eighth, tenth, twelfth, fourteenth, sixteenth and eighteenth nucleotide of the sense strand. Additionally, it is to be noted that the in case of these particular nucleic acids according to the present invention the first stretch is identical to the first strand and the second stretch is identical to the second strand and these nucleic acids are also blunt ended.

According to one preferred embodiment of the invention, the antisense and the sense strands of the siRNA molecule are both phosphorylated only at the 3′-terminus and not at the 5′-terminus. According to another preferred embodiment of the invention, the antisense and the sense strands are both non-phosphorylated both at the 3′-terminus and also at the 5′-terminus. According to yet another preferred embodiment of the invention, the 1^(st) nucleotide in the 5′ position in the sense strand is specifically modified to abolish any possibility of in vivo 5′-phosphorylation.

In another embodiment of the compound of the invention, the ribonucleotides at the 5′ and 3′ termini of the antisense strand are modified in their sugar residues, and the ribonucleotides at the 5′ and 3′ termini of the sense strand are unmodified in their sugar residues.

The invention further provides a vector capable of expressing any of the aforementioned oligoribonucleotides in unmodified form in a cell after which appropriate modification may be made.

The invention also provides a composition comprising one or more of the compounds of the invention in a carrier, preferably a pharmaceutically acceptable carrier. This composition may comprise a mixture of two or more different siRNAs.

The invention also provides a composition which comprises the above compound of the invention covalently or non-covalently bound to one or more compounds of the invention in an amount effective to inhibit Nrf2 and a carrier. This composition may be processed intracellularly by endogenous cellular complexes to produce one or more oligoribonucleotides of the invention.

The invention also provides a composition comprising a carrier and one or more of the compounds of the invention in an amount effective to down-regulate expression in a cell of Nrf2, which compound comprises a sequence substantially complementary to the sequence of (N)_(x).

Additionally the invention provides a method of down-regulating the expression of Nrf2 by at least 50% as compared to a control comprising contacting an mRNA transcript of Nrf2 with one or more of the compounds of the invention.

In one embodiment the oligoribonucleotide is down-regulating Nrf2, whereby the down-regulation of Nrf2 is selected from the group comprising down-regulation of gene function, down-regulation of polypeptide and down-regulation of mRNA expression.

In one embodiment the compound is down-regulating the Nrf2 polypeptide, whereby the down-regulation is selected from the group comprising down-regulation of function (which may be examined by an enzymatic assay or a binding assay with a known interactor of the native gene/polypeptide, inter alia), down-regulation of protein (which may be examined by Western blotting, ELISA or immuno-precipitation, inter alia) and down-regulation of mRNA expression (which may be examined by Northern blotting, quantitative RT-PCR, in-situ hybridisation or microarray hybridisation, inter alia).

The invention also provides a method of treating a patient suffering from a disease accompanied by an elevated level of Nrf2, the method comprising administering to the patient a composition of the invention in a therapeutically effective dose thereby treating the patient.

More particularly, the invention provides an oligoribonucleotide wherein one strand comprises consecutive nucleotides having, from 5′ to 3′, the sequence set forth in Table A (19mer siRNA molecules, SEQ ID NOS: 1-1006) or in Table B (21mer siRNA molecules, SEQ ID NOS: 1007-2006), or a homolog thereof wherein in up to 2 of the nucleotides in each terminal region a base is altered. Preferred 19mer siRNA molecules comprise the sense and corresponding antisense sequences listed in Table C. More preferred siRNA comprises SEQ ID NO: 10 (sense sequence) and SEQ ID NO: 513 (antisense sequence).

The terminal region of the oligonucleotide refers to bases 1-4 and/or 16-19 in the 19-mer sequence and to bases 1-4 and/or 18-21 in the 21-mer sequence.

The presently most preferred compound of the invention is a blunt-ended 19-mer oligonucleotide, i.e. x=y=19 and Z and Z′ are both absent. The oligonucleotide molecule is either phosphorylated at 3′ termini of both sense and anti-sense strands, or non-phosphorylated at all; or having I″ nucleotide in the 5′ position on the sense strand specifically modified to abolish any possibility of in vivo 5′-phosphorylation. The alternating ribonucleotides are modified at the 2′ position in both the antisense and the sense strands, wherein the moiety at the 2′ position is methoxy (2′-0-methyl) and wherein the ribonucleotides at the 5′ and 3′ termini of the antisense strand are modified in their sugar residues, and the ribonucleotides at the 5′ and 3′ termini of the sense strand are unmodified in their sugar residues.

In certain embodiments the present invention provides a compound having the structure

5′ (N)x 3′ antisense strand 3′ (N′)y 5′ sense strand wherein each of x and y=19 and (N)_(x) and (N′)_(y) are fully complementary; wherein alternating ribonucleotides in (N)x and (N′)y are modified to result in a 2′-O-methyl modification in the sugar residue of the ribonucleotides; wherein each N at the 5′ and 3′ termini of (N)_(x) are modified; wherein each N′ at the 5′ and 3′ termini of (N′)_(y) are unmodified; wherein each of (N)_(x) and (N′)_(y) is selected from the group of oligomers set forth in any one of Tables A and B (SEQ ID NOS: 1-2006). (N)_(x) and (N′)_(y) may be phosphorylated or non-phosphorylated at the 3′ and 5′ termini.

In certain embodiments of the invention, alternating ribonucleotides are modified in the 2′ position of the sugar residue in both the antisense and the sense strands of the compound. In particular the siRNAs may be modified such that a 2′-O-methyl (Me) group is present on the first, third, fifth, seventh, ninth, eleventh, thirteenth, fifteenth, seventeenth and nineteenth nucleotide of the antisense strand, whereby the very same modification, i.e. a 2′-O-Me group, is present at the second, fourth, sixth, eighth, tenth, twelfth, fourteenth, sixteenth and eighteenth nucleotide of the sense strand. These particular siRNA compounds may also be blunt ended.

In certain embodiments of the compounds of the invention having alternating ribonucleotides modified in one or both of the antisense and the sense strands of the compound; for 19-mers and 23-mers the ribonucleotides at the 5′ and 3′ termini of the antisense strand are modified in their sugar residues, and the ribonucleotides at the 5′ and 3′ termini of the sense strand are unmodified in their sugar residues. For 21-mers the ribonucleotides at the 5′ and 3′ termini of the sense strand are modified in their sugar residues, and the ribonucleotides at the 5′ and 3′ termini of the antisense strand are unmodified in their sugar residues. As mentioned above, it is preferred that the middle nucleotide of the antisense strand is unmodified.

In some preferred embodiments the middle ribonucleotide in the first strand (antisense) is an unmodified nucleotide. For example, in a 19-oligomer antisense strand, ribonucleotide number 10 is unmodified; in a 21-oligomer antisense strand, ribonucleotide number 11 is unmodified; and in a 23-oligomer antisense strand, ribonucleotide number 12 is unmodified. The modifications or pattern of modification, if any, of the siRNA must be planned to allow for this.

Delivery:

The siRNA molecules of the present invention may be delivered to the target tissue (such as the lung) by direct application of the naked molecules admixed with a carrier or a diluent using an aerosol.

For administration via the upper respiratory tract, the composition is formulated into a solution, e.g., water or isotonic saline, buffered or unbuffered, or as a suspension, at an appropriate concentration for oro-nasal administration as an aerosol. Preferably, such solutions or suspensions are isotonic relative to nasal secretions and of about the same pH, ranging e.g., from about pH 4.0 to about pH 7.4 or, from pH 6.0 to pH 7.0. Buffers should be physiologically compatible and include, simply by way of example, phosphate buffers. For example, a representative nasal decongestant is described as being buffered to a pH of about 6.2 (Remington's Pharmaceutical Sciences 16th edition, Ed. Arthur Osol, page 1445 (1980)). One skilled in the art can readily determine a suitable saline content and pH for an innocuous aqueous solution for nasal and/or upper respiratory administration.

Other suitable aqueous vehicles include, but are not limited to, Ringer's solution and isotonic sodium chloride. Aqueous suspensions may include suspending agents such as cellulose derivatives, sodium alginate, polyvinyl-pyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.

The compositions may contain minor amounts of polymers, surfactants, or other excipients well known to those of the art. In this context, “minor amounts” means no auxiliary agents or substances are present that might affect or mediate uptake of nucleic acid in the cells of the lungs.

Aerosol dosage, formulations and delivery systems may be selected for a particular therapeutic application, as described, for example, in Gonda, I. “Aerosols for delivery of therapeutic and diagnostic agents to the respiratory tract,” in Critical Reviews in Therapeutic Drug Carrier Systems, 6:273-313, 1990; and in Moren, “Aerosol dosage forms and formulations,” in: Aerosols in Medicine, Principles, Diagnosis and Therapy, Moren, et al., Eds. Elsevier, Amsterdam, 1985. The term aerosol as used herein refers to any preparation of a fine mist of particles, which can be in solution or a suspension, whether or not it is produced using a propellant. Aerosols can be produced using standard techniques, such as ultrasonication or high pressure treatment.

Various suitable devices and methods of inhalation which can be used to administer particles to a patient's respiratory tract are known in the art. Nebulizers create a fine mist from a solution or suspension, which is inhaled by the patient. The devices described in U.S. Pat. No. 5,709,202 to Lloyd, et al., can be used. An MDI typically includes a pressurized canister having a meter valve, wherein the canister is filled with the solution or suspension and a propellant. The solvent itself may function as the propellant, or the composition may be combined with a propellant, such as freon. The composition is a fine mist when released from the canister due to the release in pressure. The propellant and solvent may wholly or partially evaporate due to the decrease in pressure.

The term “naked siRNA” refers to siRNA molecules that are free from any delivery vehicle that acts to assist, promote or facilitate entry into the cell, including viral sequences, viral particles, liposome formulations, lipofectin or precipitating agents and the like. For example, siRNA in PBS is “naked siRNA”. However, the siRNA molecules of the invention can also be delivered in liposome formulations and lipofectin formulations and the like and can be prepared by methods well known to those skilled in the art. Such methods are described, for example, in U.S. Pat. Nos. 5,593,972, 5,589,466, and 5,580,859, which are herein incorporated by reference.

Delivery systems aimed specifically at the enhanced and improved delivery of siRNA into mammalian cells have been developed, see, for example, Shen et al (FEBS letters 539: 111-114 (2003)), Xia et al., Nature Biotechnology 20: 1006-1010 (2002), Reich et al., Molecular Vision 9: 210-216 (2003), Sorensen et al. (J. Mol. Biol. 327: 761-766 (2003), Lewis et al., Nature Genetics 32: 107-108 (2002) and Simeoni et al., Nucleic Acids Research 31, 11: 2717-2724 (2003). siRNA has recently been successfully used for inhibition in primates; for further details see Tolentino et al., Retina 24(1) February 2004 I 132-138. Respiratory formulations for siRNA are described in U.S. patent application No. 2004/0063654 of Davis et al. Cholesterol-conjugated siRNAs (and other steroid and lipid conjugated siRNAs) can been used for delivery see Soutschek et al Nature 432: 173-177 (2004); and Lorenz et al. Bioorg. Med. Chemistry. Lett. 14:4975-4977 (2004).

The siRNAs or pharmaceutical compositions of the present invention are administered and dosed in accordance with good medical practice, taking into account the clinical condition of the individual patient, the disease to be treated, the site and method of administration, scheduling of administration, patient age, sex, body weight and other factors known to medical practitioners.

The “therapeutically effective dose” for purposes herein is thus determined by such considerations as are known in the art. The dose must be effective to achieve improvement including but not limited to improved survival rate or more rapid recovery, or improvement or elimination of symptoms and other indicators as are selected as appropriate measures by those skilled in the art. The compounds of the present invention can be administered by any of the conventional routes of administration. It should be noted that the compound can be administered as the compound or as pharmaceutically acceptable salt and can be administered alone or as an active ingredient in combination with pharmaceutically acceptable carriers, solvents, diluents, excipients, adjuvants and vehicles. The compounds can be administered orally, subcutaneously or parenterally including intravenous, intraarterial, intramuscular, intraperitoneally, and intranasal administration as well as intrathecal and infusion techniques. Implants of the compounds are also useful. Liquid forms may be prepared for injection, the term including subcutaneous, transdermal, intravenous, intramuscular, intrathecal, and other parental routes of administration. The liquid compositions include aqueous solutions, with and without organic co-solvents, aqueous or oil suspensions, emulsions with edible oils, as well as similar pharmaceutical vehicles. In addition, under certain circumstances the compositions for use in the novel treatments of the present invention may be formed as aerosols, for intranasal and like administration. The patient being treated is a warm-blooded animal and, in particular, mammals including man. The pharmaceutically acceptable carriers, solvents, diluents, excipients, adjuvants and vehicles as well as implant carriers generally refer to inert, non-toxic solid or liquid fillers, diluents or encapsulating material not reacting with the active ingredients of the invention and they include liposomes and microspheres.

Examples of delivery systems useful in the present invention include U.S. Pat. Nos. 5,225,182; 5,169,383; 5,167,616; 4,959,217; 4,925,678; 4,487,603; 4,486,194; 4,447,233; 4,447,224; 4,439,196; and 4,475,196. Many other such implants, delivery systems, and modules are well known to those skilled in the art. In one specific embodiment of this invention topical and transdermal formulations are particularly preferred.

In general, the active dose of compound for humans is in the range of from 1 ng/kg to about 20-100 mg/kg body weight per day, preferably about 0.01 mg to about 2-10 mg/kg body weight per day, in a regimen of one dose per day or twice or three or more times per day for a period of 1-4 weeks or longer.

The term “treatment” as used herein refers to administration of a therapeutic substance effective to ameliorate symptoms associated with a disease, to lessen the severity or cure the disease, or to prevent the disease from occurring. Thus “treatment” refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) a disease in particular a cancerous disease such as lung cancer, breast cancer, cervical cancer, colon cancer, gastric cancer, kidney cancer, leukemia, liver cancer, lymphoma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, sarcoma, skin cancer, testicular cancer, and uterine cancer. In a particular embodiment, the cancer is lung cancer such as non-small-cell lung carcinoma (NSCLC) or small-cell lung carcinoma

In another aspect of the invention a pharmaceutical composition is provided which comprises any of the above oligoribonucleotides or vectors and a pharmaceutically acceptable carrier. Another aspect of the invention is the use of a therapeutically effective amount of any of the above oligoribonucleotides or vectors for the preparation of a medicament for treating a patient suffering from a cancer disease.

By “cancerous disease” is meant any disease that is caused by or results in inappropriately high levels of cell division, inappropriately low levels of apoptosis, or both. Examples of cancerous diseases include, without limitation, leukemias (e.g., acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (Hodgkin's disease, non-Hodgkin's disease), Waldenstrom's macroglobulinemia, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangio sarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyo sarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, nile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, crailiopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodenroglioma, schwamioma, meningioma, melanoma, neuroblastoma, and retinoblastoma).

In one embodiment, the present invention provides a pharmaceutical composition comprising an Nrf2 inhibitory molecule, preferable an siRNA molecule that decreases the expression of the Nrf2 gene in combination with a chemotherapeutic agent. Without being bound by theory, since Nrf2 positively regulates drug detoxification enzymes, targeting this molecule may have a broad effect on all anticancer drugs. In various embodiments, the Nrf2 inhibitory molecule is administered prior to, concurrently with, or following administration of a chemotherapeutic. Without wishing to be bound by theory, administration of an Nrf2 inhibitory molecule likely enhances the accumulation or efficacy of a chemotherapeutic agent.

Compositions and methods of the invention may be used in combination with any conventional therapy known in the art. In one embodiment, the Nrf2 inhibitory molecules of the invention may be used in combination with one or more anti-cancerous therapy known in the art. Thus the Nrf2 inhibitors may be used before, during or following anti-cancerous therapy Exemplary anti-cancerous therapies include, for example, chemotherapy, cryotherapy, hormone therapy, growth factor inhibitors, radiotherapy, and surgery. Chemotherapeutics typically used in the treatment of a cancerous disease are for example abiraterone acetate, altretamine, anhydrovinbiastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide, bleomycin, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proly-1-Lproline-t-butylamide, cachectin, cemadotin, chiorambucil, cyclophosphamide, 3′,4′-didehydro-4′-deoxy-8′-norvin-caleukoblastine, docetaxol, doxetaxel, cyclophosphamide, carboplatin, carmustine (BCNTJ), cisplatin, cryptophycin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, daunorubicin, dolastatin, doxorubicin (adriamycin), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea and hydroxyureataxanes, ifosfamide, liarozole, lonidamine, lomustine (CCNU), mechlorethamine (nitrogen mustard), melphalan, mivobulin isethionate, rhizoxin, sertenef, streptozocin, mitomycin, methotrexate, 5-fluorouracil, nilutamide, onapristone, oxaliplatin paclitaxel, prednimustine, procarbazine, RPR1 09881, satrapaltin, stramustine phosphate, tamoxifen, tasonermin, taxol, tretinoin, vinbiastine, vincristine, vindesine sulfate, and vinflunine. Other examples of chemotherapeutic agents can be found in Cancer Principles and Practice of Oncology by V. T. Devita and S. Heliman (editors), 6th edition (Fel. 15, 2001), Lippincott Williams & Wilkins Publishers. A preferred combination according to the present invention is an siRNA molecule targeting Nrf2 with platinum drugs, more preferably an siRNA molecule set forth in Table C with a platinum drug, preferably carboplatin. Without being bound by theory, the Nrf2 siRNA inhibitors of the invention are found to be efficient promoters for the antineoplastic potential of platinum drugs, causing additive/synergistic effects in cancer cells. Platinum drugs include carboplatin, cisplatin, oxaliplatin and satrapaltin inter alia; see Kelland and Farrell, Platinum-based drugs in cancer therapy (Cancer drug discovery & development) Lavoisier 2000 which is hereby incorporated by reference.

The compounds which reduce or prevent the cancerous disease such as lung cancer, eg the novel siRNAs inter alia are preferably administered directly to the inner lung as naked siRNA in a vehicle such as PBS or other physiological solutions, but may alternatively be administered with a delivery vehicle as described above.

The present invention also provides for a process of preparing a pharmaceutical composition, which comprises:

-   -   obtaining one or more double stranded compound of the invention;         and     -   admixing said compound with a pharmaceutically acceptable         carrier.

The present invention also provides for a process of preparing a pharmaceutical composition, which comprises admixing one or more compounds of the present invention with a pharmaceutically acceptable carrier.

In a preferred embodiment, the compound used in the preparation of a pharmaceutical composition is admixed with a carrier in a pharmaceutically effective dose. In a particular embodiment the compound of the present invention is conjugated to a steroid or to a lipid or to another suitable molecule e.g. to cholesterol.

Modifications or analogs of nucleotides can be introduced to improve the therapeutic properties of the nucleotides. Improved properties include increased nuclease resistance and/or increased ability to permeate cell membranes.

Accordingly, the present invention also includes all analogs of, or modifications to, a oligonucleotide of the invention that does not substantially affect the function of the polynucleotide or oligonucleotide. In a preferred embodiment such modification is related to the base moiety of the nucleotide, to the sugar moiety of the nucleotide and/or to the phosphate moiety of the nucleotide.

In embodiments of the invention, the nucleotides can be selected from naturally occurring or synthetically modified bases. Naturally occurring bases include adenine, guanine, cytosine, thymine and uracil. Modified bases of the oligonucleotides include inosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl-, 2-propyl- and other alkyl-adenines, 5-halo uracil, 5-halo cytosine, 6-aza cytosine and 6-aza thymine, pseudo uracil, 4-thiuracil, 8-halo adenine, 8-aminoadenine, 8-thiol adenine, 8-thiolalkyl adenine, 8-hydroxyl adenine and other 8-substituted adenines, 8-halo guanine, 8-amino guanine, 8-thiol guanine, 8-thioalkyl guanine, 8-hydroxyl guanine and other substituted guanines, other aza and deaza adenines, other aza and deaza guanines, 5-trifluoromethyl uracil and 5-trifluoro cytosine.

In addition, analogs of nucleotides can be prepared wherein the structures of the nucleotides are fundamentally altered and are better suited as therapeutic or experimental reagents. An example of a nucleotide analog is a peptide nucleic acid (PNA) wherein the deoxyribose (or ribose) phosphate backbone in DNA (or RNA) is replaced with a polyamide backbone similar to that found in peptides. PNA analogs have been shown to be resistant to degradation by enzymes and to have extended lives in vivo and in vitro. Further, PNAs have been shown to bind more strongly to a complementary DNA sequence than to a DNA molecule. This observation is attributed to the lack of charge repulsion between the PNA strand and the DNA strand. Other modifications that can be made to oligonucleotides include polymer backbones, cyclic backbones, or acyclic backbones.

In one embodiment the modification is a modification of the phosphate moiety, whereby the modified phosphate moiety is selected from the group comprising phosphothioate.

The compounds of the present invention can be synthesized by any of the methods that are well-known in the art for synthesis of ribonucleic (or deoxyribonucleic) oligonucleotides. Such synthesis is, among others, described in Beaucage S. L. and Iyer R. P., Tetrahedron 1992; 48: 2223-2311, Beaucage S. L. and Iyer R. P., Tetrahedron 1993; 49: 6123-6194 and Caruthers M. H. et. al., Methods Enzymol. 1987; 154: 287-313; the synthesis of thioates is, among others, described in Eckstein F., Annu. Rev. Biochem. 1985; 54: 367-402, the synthesis of RNA molecules is described in Sproat B., in Humana Press 2005 edited by Herdewijn P.; Kap. 2: 17-31 and respective downstream processes are, among others, described in Pingoud A. et. al., in IRL Press 1989 edited by Oliver R. W. A.; Kap. 7: 183-208 and Sproat B., in Humana Press 2005 edited by Herdewijn P.; Kap. 2: 17-31 (supra).

Other synthetic procedures are known in the art e.g. the procedures as described in Usman et al., 1987, J. Am. Chem. Soc., 109, 7845; Scaringe et al., 1990, Nucleic Acids Res., 18, 5433; Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684; and Wincott et al., 1997, Methods Mol. Bio., 74, 59, and these procedures may make use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. The modified (e.g. 2′-O-methylated) nucleotides and unmodified nucleotides are incorporated as desired.

The oligonucleotides of the present invention can be synthesized separately and joined together post-synthetically, for example, by ligation (Moore et al., 1992, Science 256, 9923; Draper et al., International PCT publication No. WO93/23569; Shabarova et al., 1991, Nucleic Acids Research 19, 4247; Bellon et al., 1997, Nucleosides & Nucleotides, 16, 951; Bellon et al., 1997, Bioconjugate Chem. 8, 204), or by hybridization following synthesis and/or deprotection.

It is noted that a commercially available machine (available, inter alia, from Applied Biosystems) can be used; the oligonucleotides are prepared according to the sequences disclosed herein. Overlapping pairs of chemically synthesized fragments can be ligated using methods well known in the art (e.g., see U.S. Pat. No. 6,121,426). The strands are synthesized separately and then are annealed to each other in the tube. Then, the double-stranded siRNAs are separated from the single-stranded oligonucleotides that were not annealed (e.g. because of the excess of one of them) by HPLC. In relation to the siRNAs or siRNA fragments of the present invention, two or more such sequences can be synthesized and linked together for use in the present invention.

The compounds of the invention can also be synthesized via a tandem synthesis methodology, as described in US patent application publication No. US2004/0019001 (McSwiggen), wherein both siRNA strands are synthesized as a single contiguous oligonucleotide fragment or strand separated by a cleavable linker which is subsequently cleaved to provide separate siRNA fragments or strands that hybridize and permit purification of the siRNA duplex. The linker can be a polynucleotide linker or a non-nucleotide linker.

The present invention further provides for a pharmaceutical composition comprising two or more siRNA molecules for the treatment of any of the diseases and conditions mentioned herein, whereby said two molecules may be physically mixed together in the pharmaceutical composition in amounts which generate equal or otherwise beneficial activity, or may be covalently or non-covalently bound, or joined together by a nucleic acid linker of a length ranging from 2-100, preferably 2-50 or 2-30 nucleotides. In one embodiment, the siRNA molecules are comprised of a double-stranded nucleic acid structure as described herein, wherein the two siRNA sequences are selected from Tables A or B.

In another embodiment, the siRNA molecules are comprised of a double-stranded nucleic acid structure, wherein the first siRNA sequence is selected from Tables A or B, and the second siRNA molecule targets another anti-cancer gene, thereby providing beneficial activity. The tandem double-stranded structure which comprises two or more siRNA sequences is processed intracellularly to form two or more different siRNAs. Such second siRNA molecule is preferably an siRNA molecule that targets a anti-cancer gene. Preferred anti-cancer genes are for example gene encoding growth factors such as insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) and platelet derived growth factor (PDGF).

The siRNA molecules are covalently or non-covalently bound or joined by a linker to form a tandem siRNA molecule. Such tandem siRNA molecules comprising two siRNA sequences are typically of 38-150 nucleotides in length, more preferably 38 or 40-60 nucleotides in length, and longer accordingly if more than two siRNA sequences are included in the tandem molecule. A longer tandem molecule comprised of two or more longer sequences which encode siRNA produced via internal cellular processing, e.g., long dsRNAs, is also envisaged, as is a tandem molecule encoding two or more shRNAs. Such tandem molecules are also considered to be a part of the present invention.

siRNA molecules that target Nrf2 may be the main active component in a pharmaceutical composition, or may be one active component of a pharmaceutical composition containing two or more siRNAs (or molecules which encode or endogenously produce two or more siRNAs, be it a mixture of molecules or one or more tandem molecules which encode two or more siRNAs), said pharmaceutical composition further being comprised of one or more additional siRNA molecule which targets one or more additional gene. Simultaneous inhibition of Nrf2 and said additional gene(s) will likely have an additive or synergistic effect for treatment of the diseases disclosed herein.

In a preferred embodiment, the one or more additional siRNA molecules target an anti-cancer gene, thus having an additive or synergistic effect with the Nrf2 siRNA. The additional siRNA molecules may target one or more of the anti-cancer genes defined above.

As disclosed herein, aptamers may also be used in the present invention alone or in combination with the novel siRNAs disclosed herein for targeting NRF2 and for the treatment of any one of the conditions disclosed herein. For example, an aptamer can be used with any one of the siRNAs disclosed herein in combination therapy for the treatment of any one of the conditions disclosed herein. The novel pharmaceutical composition employed for such a combination therapy, which is also part of the present invention, may comprise an siRNA of the present invention covalently or non-covalently attached to an aptamer. Aptamers are RNA or DNA single-strand or double-strand oligonucleic acids which bind to a target protein and do not generally exhibit non-specific effects. Aptamers can be modified for stability or other desired qualities in accordance with any nucleic acid modifications disclosed herein and/or known to one of skill in the art. Modifications to aptamers can be introduced anywhere in the molecule, such as the 5′ or 3′ termini, or at any internally defined modification site. For example, RNA aptamers can be stabilized with 2′-fluoro or 2′-amino modified pyrimidines. Aptamers can also be linked to reporter molecules or linker chemistries and can be attached to beads or other solid support if necessary (e.g., 5′ or 3′ amino, thiol ester or biotin groups). Thioaptamers are aptamers which contain sulfur modifications at specific internucleoside phosphoryl sites, and may possess enhanced stability, nuclease resistance, target affinity and/or selectivity. Examples of thioaptamers include phosphoromonothioate (S-ODN) and phosphorodithioate (S2-ODN) oligodeoxy thioaptamers. For further information on aptamers and thioaptamers see U.S. Pat. Nos. 5,218,088 and 6,423,493.

Additionally, the siRNA disclosed herein or any nucleic acid molecule comprising or encoding such siRNA can be linked or bound (covalently or non-covalently) to antibodies (including aptamer molecules) against cell surface internalizable molecules expressed on the target cells, in order to achieve enhanced targeting for treatment of the diseases disclosed herein. For example, anti-Fas antibody (preferably a neutralizing antibody) may be combined (covalently or non-covalently) with a Nrf2 siRNA molecule. In another example, an aptamer which can act like a ligand/antibody may be combined (covalently or non-covalently) with a Nrf2 siRNA molecule.

The term “Covalent bonding” as used herein refers to chemical bonding that is characterized by the sharing of pairs of electrons between atoms.

The term “Noncovalent bonding” as used herein refers to a variety of interactions that are not covalent in nature between molecules or parts of molecules that provide force to hold the molecules or parts of molecules together, usually in a specific orientation or conformation. These noncovalent interactions include: ionic bonds, hydrophobic interactions, hydrogen bonds, Van der Waals forces anddipole-dipole bonds.

The compounds of the present invention can be delivered either directly or with viral or non-viral vectors. When delivered directly the sequences are generally rendered nuclease resistant. Alternatively the sequences can be incorporated into expression cassettes or constructs such that the sequence is expressed in the cell as discussed herein below. Generally the construct contains the proper regulatory sequence or promoter to allow the sequence to be expressed in the targeted cell. Vectors optionally used for delivery of the compounds of the present invention are commercially available, and may be modified for the purpose of delivery of the compounds of the present invention by methods known to one of skill in the art.

It is also envisaged that a long oligonucleotide (typically 25-500 nucleotides in length) comprising one or more stem and loop structures, where stern regions comprise the sequences of the oligonucleotides of the invention, may be delivered in a carrier, preferably a pharmaceutically acceptable carrier, and may be processed intracellularly by endogenous cellular complexes (e.g. by DROSHA and DICER as described above) to produce one or more smaller double stranded oligonucleotides (siRNAs) which are oligonucleotides of the invention. This oligonucleotide can be termed a tandem shRNA construct. It is envisaged that this long oligonucleotide is a single stranded oligonucleotide comprising one or more stem and loop structures, wherein each stem region comprises a sense and corresponding antisense siRNA sequence of an NRF2 gene. In particular, it is envisaged that this oligonucleotide comprises sense and antisense siRNA sequences as depicted as any one of SEQ ID NO: 1-2006.

As used herein, the term “inhibition” of the Nrf2 gene means inhibition of the gene expression (transcription or translation) or polypeptide activity.

Although the inhibitor may be an siRNA molecule, other inhibitors contemplated to be used in the methods of the invention to inhibit the Nrf2 gene and to treat the diseases and conditions described herein are inter alia antibodies, preferably neutralizing antibodies or fragments thereof, including single chain antibodies, antisense oligonucleotides, antisense DNA or RNA molecules, aptamers, proteins, polypeptides and peptides including peptidomimetics and dominant negatives, and also expression vectors expressing all the above. Additional inhibitors may be small chemical molecules, which generally have a molecular weight of less than 2000 daltons, more preferably less than 1000 daltons, even more preferably less than 500 daltons. These inhibitors may act as follows: small molecules may affect expression and/or activity; antibodies may affect activity; all kinds of antisense may affect the gene expression; and dominant negative polypeptides and peptidomimetics may affect activity; expression vectors may be used inter alia for delivery of antisense or dominant-negative polypeptides or antibodies.

The term “antibody” refers to IgG, IgM, IgD, IgA, and IgE antibody, inter alia. The definition includes polyclonal antibodies or monoclonal antibodies. This term refers to whole antibodies or fragments of antibodies comprising an antigen-binding domain, e.g. antibodies without the Fc portion, single chain antibodies, miniantibodies, fragments consisting of essentially only the variable, antigen-binding domain of the antibody, etc. The term “antibody” may also refer to antibodies against polynucleotide sequences obtained by cDNA vaccination. The term also encompasses antibody fragments which retain the ability to selectively bind with their antigen or receptor and are exemplified as follows, inter alia:

(1) Fab, the fragment which contains a monovalent antigen-binding fragment of an antibody molecule which can be produced by digestion of whole antibody with the enzyme papain to yield a light chain and a portion of the heavy chain;

(2) (Fab′)₂, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab′₂) is a dimer of two Fab fragments held together by two disulfide bonds;

(3) Fv, defined as a genetically engineered fragment containing the variable region of the light chain and the variable region of the heavy chain expressed as two chains; and

(4) Single chain antibody (SCA), defined as a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain linked by a suitable polypeptide linker as a genetically fused single chain molecule.

Antisense Molecules

By the term “antisense” (AS) or “antisense fragment” is meant a polynucleotide fragment (comprising either deoxyribonucleotides, ribonucleotides or a mixture of both) having inhibitory antisense activity, said activity causing a decrease in the expression of the endogenous genomic copy of the corresponding gene. An AS polynucleotide is a polynucleotide which comprises consecutive nucleotides having a sequence of sufficient length and homology to a sequence present within the sequence of the target gene to permit hybridization of the AS to the gene. Many reviews have covered the main aspects of antisense (AS) technology and its enormous therapeutic potential (Aboul-Fadl T., Curr Med. Chem. 2005; 12(19):2193-214; Crooke S T, Curr Mol. Med. 2004 August; 4(5):465-87; Crooke S T, Annu Rev Med. 2004; 55:61-95; Vacek M et al., Cell Mol Life Sci. 2003 May; 60(5):825-33; Cho-Chung Y S, Arch Pharm Res. 2003 March; 26(3):183-91. There are further reviews on the chemical (Crooke, 1995; Uhlmann et al, 1990), cellular (Wagner, 1994) and therapeutic (Hanania, et al, 1995; Scanlon, et al, 1995; Gewirtz, 1993) aspects of this technology. Antisense intervention in the expression of specific genes can be achieved by the use of synthetic AS oligonucleotide sequences (for recent reports see Lefebvre-d'Hellencourt et al, 1995; Agrawal, 1996; LevLehman et al, 1997).

AS oligonucleotide sequences may be short sequences of DNA, typically 15-30 mer but may be as small as 7 mer (Wagner et al, 1996), designed to complement a target mRNA of interest and form an RNA:AS duplex. This duplex formation can prevent processing, splicing, transport or translation of the relevant in RNA. Moreover, certain AS nucleotide sequences can elicit cellular RNase H activity when hybridized with their target mRNA, resulting in mRNA degradation (Calabretta et al, 1996 Semin Oncol. 23(1):78-87). In that case, RNase H will cleave the RNA component of the duplex and can potentially release the AS to further hybridize with additional molecules of the target RNA. An additional mode of action results from the interaction of AS with genomic DNA to form a triple helix which can be transcriptionally inactive.

The sequence target segment for the antisense oligonucleotide is selected such that the sequence exhibits suitable energy related characteristics important for oligonucleotide duplex formation with their complementary templates, and shows a low potential for self-dimerization or self-complementation [Anazodo et al., 19961. For example, the computer program OLIGO (Primer Analysis Software, Version 3.4), can be used to determine antisense sequence melting temperature, free energy properties, and to estimate potential self-dimer formation and self-complimentary properties. The program allows the determination of a qualitative estimation of these two parameters (potential self-dimer formation and self-complimentary) and provides an indication of “no potential” or “some potential” or “essentially complete potential”. Using this program target segments are generally selected that have estimates of no potential in these parameters. However, segments can be used that have “some potential” in one of the categories. A balance of the parameters is used in the selection as is known in the art. Further, the oligonucleotides are also selected as needed so that analogue substitution do not substantially affect function.

Phosphorothioate antisense oligonucleotides do not normally show significant toxicity at concentrations that are effective and exhibit sufficient pharmacodynamic half-lives in animals (Agarwal et al., 1996) and are nuclease resistant. Antisense induced loss-of-function phenotypes related with cellular development were shown for the glial fibrillary acidic protein (GFAP), for the establishment of tectal plate formation in chick (Galileo et al., 1991) and for the N-myc protein, responsible for the maintenance of cellular heterogeneity in neuroectodermal cultures (ephithelial vs. neuroblastic cells, which differ in their colony forming abilities, tumorigenicity and adherence) (Rosolen et al., 1990; Whitesell et al, 1991). Antisense oligonucleotide inhibition of basic fibroblast growth factor (bFgF), having mitogenic and angiogenic properties, suppressed 80% of growth in glioma cells (Morrison, 1991) in a saturable and specific manner. Being hydrophobic, antisense oligonucleotides interact well with phospholipid membranes (Akhter et al., 1991). Following their interaction with the cellular plasma membrane, they are actively (or passively) transported into living cells (Loke et al., 1989), in a saturable mechanism predicted to involve specific receptors (Yakubov et al., 1989).

Ribozymes

A “ribozyme” is an RNA molecule that possesses RNA catalytic ability (see Cech for review) and cleaves a specific site in a target RNA. In accordance with the present invention, ribozymes which cleave mRNA may be utilized as inhibitors. This may be necessary in cases where antisense therapy is limited by stoichiometric considerations (Sarver et al., 1990, Gene Regulation and Aids, pp. 305-325). Ribozymes can then be used that will target the a gene associated with a bone marrow disease. The number of RNA molecules that are cleaved by a ribozyme is greater than the number predicted by stochiochemistry. (Hampel and Tritz, 1989; Uhlenbeck, 1987).

Ribozymes catalyze the phosphodiester bond cleavage of RNA. Several ribozyme structural families have been identified including Group I introns, RNase P, the hepatitis delta virus ribozyme, hammerhead ribozymes and the hairpin ribozyme originally derived from the negative strand of the tobacco ringspot virus satellite RNA (sTRSV) (Sullivan, 1994; U.S. Pat. No. 5,225,347). The latter two families are derived from viroids and virusoids, in which the ribozyme is believed to separate monomers from oligomers created during rolling circle replication (Symons, 1989 and 1992). Hammerhead and hairpin ribozyme motifs are most commonly adapted for trans-cleavage of mRNAs for gene therapy (Sullivan, 1994). In general the ribozyme has a length of from about 30-100 nucleotides. Delivery of ribozymes is similar to that of AS fragments and/or siRNA molecules.

Screening of Inactivation Compounds for Nrf2:

Some of the compounds and compositions of the present invention may be used in a screening assay for identifying and isolating compounds that modulate the activity of the NRF2 gene, in particular compounds that modulate a disorder accompanied by an elevated level of NRF2. The compounds to be screened comprise inter alia substances such as small chemical molecules and antisense oligonucleotides.

The inhibitory activity of the compounds of the present invention on Nrf2 expression may be used to determine the interaction of an additional compound with the target polypeptide, e.g., if the additional compound competes with the oligonucleotides of the present invention for inhibition of Nrf2, or if the additional compound rescues said inhibition. The inhibition or activation can be tested by various means, such as, inter alia, assaying for the product of the activity of the Nrf2 polypeptide or displacement of binding compound from the Nrf2 polypeptide in radioactive or fluorescent competition assays.

The present invention is illustrated in detail below with reference to the Examples, but is not to be construed as being limited thereto.

Citation of any document herein is not intended as an admission that such document is pertinent prior art, or considered material to the patentability of any claim of the present application. Any statement as to content or a date of any document is based on the information available to applicant at the time of filing and does not constitute an admission as to the correctness of such a statement.

EXAMPLES General Methods in Molecular Biology

Standard molecular biology techniques known in the art and not specifically described were generally followed as in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York (1989), and as in Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore, Md. (1989) and as in Perbal, A Practical Guide to Molecular Cloning, John Wiley & Sons, New York (1988), and as in Watson et al., Recombinant DNA, Scientific American Books, New York and in Birren et al (eds) Genome Analysis: A Laboratory Manual Series, Vols. 1-4 Cold Spring Harbor Laboratory Press, New York (1998) and methodology as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057 and incorporated herein by reference. Polymerase chain reaction (PCR) was carried out generally as in PCR Protocols: A Guide To Methods And Applications, Academic Press, San Diego, Calif. (1990). In situ (In cell) PCR in combination with Flow Cytometry can be used for detection of cells containing specific DNA and mRNA sequences (Testoni et al., 1996, Blood 87:3822.) Methods of performing RT-PCR are also well known in the art.

Example 1 Generation of Sequences for Active siRNA Compounds

Using proprietary algorithms and the known sequence of Nrf2 (gi2014957), the sequences of many potential siRNAs were generated. Table A shows a list of 19-mers siRNAs specific to Nrf2 which are either human-specific, or human and cross-species with other species. Table B shows a list of 21-mers siRNAs specific to Nrf2 which are either human-specific, or human and cross-species with other species. All siRNAs are depicted in 5′ to 3′ orientation, and the sense and complementary antisense sequences are depicted on the same line in the Table. The sense siRNAs in Table A have SEQ ID NOS: 1-503 and the antisense siRNAs in Table A have SEQ ID NOS: 504-1006. The sense siRNAs in Table B have SEQ ID NOS: 1007-1506 and the antisense siRNAs in Table B have SEQ ID NOS: 1507-2006.

TABLE A 19-mers siRNAs specific to Nrf2 (SEQ ID NOS: 1-1006) SEQ ID  SEQ ID NO. Sense siRNA NO. Antisense siRNA Human-20149575 1 GGGGUAAGAAUAAAGUGGC 504 GCCACUUUAUUCUUACCCC [1624-1642](19/19) 2 GGAGGGGUAAGAAUAAAGU 505 ACUUUAUUCUUACCCCUCC [1621-1639](19/19) 3 GCAAAACUAACCACUAUGU 506 ACAUAGUGGUUAGUUUUGC [2270-2288](19/19) 4 AAGGAGAAAAUGACAAAAG 507 CUUUUGUCAUUUUCUCCUU [1741-1759](19/19) 5 CAGAAUUGCAGAAAAAGAA 508 UUCUUUUUCUGCAAUUCUG [1647-1665](19/19) 6 GGGUAAGAAUAAAGUGGCU 509 AGCCACUUUAUUCUUACCC [1625-1643](19/19) 7 GCCCUCACCUGCUACUUUA 510 UAAAGUAGCAGGUGAGGGC [1010-1028](19/19) 8 AGGGGUAAGAAUAAAGUGG 511 CCACUUUAUUCUUACCCCU [1623-1641](19/19) 9 AGGAGGGGUAAGAAUAAAG 512 CUUUAUUCUUACCCCUCCU [1620-1638](19/19) 10 UCCCGUUUGUAGAUGACAA 513 UUGUCAUCUACAAACGGGA [493-511](19/19) 11 AGGAGAAAAUGACAAAAGC 514 GCUUUUGUCAUUUUCUCCU [1742-1760](19/19) 12 GCUCAGAAUUGCAGAAAAA 515 UUUUUCUGCAAUUCUGAGC [1644-1662](19/19) 13 GAGAAAGAAUUGCCUGUAA 516 UUACAGGCAAUUCUUUCUC [1392-1410](19/19) 14 UCCGGCAUUUCACUAAACA 517 UGUUUAGUGAAAUGCCGGA [1143-1161](19/19) 15 GAGUUACAGUGUCUUAAUA 518 UAUUAAGACACUGUAACUC [699-717](19/19) 16 GCCAAAACUAGUAUAGAAA 519 UUUCUAUACUAGUUUUGGC [2044-2062](19/19) 17 AGCCAGAUGUUAAGAAAAA 520 UUUUUCUUAACAUCUGGCU [1909-1927](19/19) 18 GAAGCCAGAUGUUAAGAAA 521 UUUCUUAACAUCUGGCUUC [1907-1925](19/19) 19 AGAAGCCAGAUGUUAAGAA 522 UUCUUAACAUCUGGCUUCU [1906-1924](19/19) 20 UCAGAAUUGCAGAAAAAGA 523 UCUUUUUCUGCAAUUCUGA [1646-1664](19/19) 21 UAGGAGGGGUAAGAAUAAA 524 UUUAUUCUUACCCCUCCUA [1619-1637](19/19) 22 CACUCUCUGAACUUCUAAA 525 UUUAGAAGUUCAGAGAGUG [1036-1054](19/19) 23 UAGCCCCUGUUGAUUUAGA 526 UCUAAAUCAACAGGGGCUA [628-646](19/19) 24 GAUCUGCCAACUACUCCCA 527 UGGGAGUAGUUGGCAGAUC [406-424](19/19) 25 GCAAAAUCAUAGCCAAAAC 528 GUUUUGGCUAUGAUUUUGC [2033-2051](19/19) 26 GUAAGAAGCCAGAUGUUAA 529 UUAACAUCUGGCUUCUUAC [1903-1921](19/19) 27 UAAAGUGGCUGCUCAGAAU 530 AUUCUGAGCAGCCACUUUA [1634-1652](19/19) 28 AGAAUAAAGUGGCUGCUCA 531 UGAGCAGCCACUUUAUUCU [1630-1648](19/19) 29 AGGAAAGACAAGAACAACU 532 AGUUGUUCUUGUCUUUCCU [280-298](19/19) 30 AAAGGAAAGACAAGAACAA 533 UUGUUCUUGUCUUUCCUUU [278-296](19/19) 31 AACUUGAAAAGGAAAGACA 534 UGUCUUUCCUUUUCAAGUU [271-289](19/19) 32 GACAAACAUUCAAGCCGCU 535 AGCGGCUUGAAUGUUUGUC [1443-1461](19/19) 33 GACUCCGGCAUUUCACUAA 536 UUAGUGAAAUGCCGGAGUC [1140-1158](19/19) 34 ACCAAAACCACCCUGAAAG 537 CUUUCAGGGUGGUUUUGGU [1099-1117](19/19) 35 ACUCUCUGAACUUCUAAAU 538 AUUUAGAAGUUCAGAGAGU [1037-1055](19/19) 36 GACAGUGAACUCAUUAAAU 539 AUUUAAUGAGUUCACUGUC [911-929](19/19) 37 AAGUUUGGGAGGAGCUAUU 540 AAUAGCUCCUCCCAAACUU [670-688](19/19) 38 UUGUAGAUGACAAUGAGGU 541 ACCUCAUUGUCAUCUACAA [499-517](19/19) 39 AAACUAACCACUAUGUACU 542 AGUACAUAGUGGUUAGUUU [2273-2291](19/19) 40 CAAAAUCAUAGCCAAAACU 543 AGUUUUGGCUAUGAUUUUG [2034-2052](19/19) 41 UAAGAAGCCAGAUGUUAAG 544 CUUAACAUCUGGCUUCUUA [1904-1922](19/19) 42 ACGUGAUGAAGAUGGAAAA 545 UUUUCCAUCUUCAUCACGU [1817-1835](19/19) 43 CUGGAAAAUAUAGUAGAAC 546 GUUCUACUAUAUUUUCCAG [1668-1686](19/19) 44 GAAAAGGAAAGACAAGAAC 547 GUUCUUGUCUUUCCUUUUC [276-294](19/19) 45 UGAAAAGGAAAGACAAGAA 548 UUCUUGUCUUUCCUUUUCA [275-293](19/19) 46 UGGAGACACACUACUUGGC 549 GCCAAGUAGUGUGUCUCCA [1211-1229](19/19) 47 UCAACCAAAACCACCCUGA 550 UCAGGGUGGUUUUGGUUGA [1096-1114](19/19) 48 CCCUCACCUGCUACUUUAA 551 UUAAAGUAGCAGGUGAGGG [1011-1029](19/19) 49 UGACAGUGAACUCAUUAAA 552 UUUAAUGAGUUCACUGUCA [910-928](19/19) 50 UUGGAGGCAAGAUAUAGAU 553 AUCUAUAUCUUGCCUCCAA [182-200](19/19) 51 GAGCUAUUAUCCAUUCCUG 554 CAGGAAUGGAUAAUAGCUC [681-699](19/19) 52 UAGAUGACAAUGAGGUUUC 555 GAAACCUCAUUGUCAUCUA [502-520](19/19) 53 UGCCCACAUUCCCAAAUCA 556 UGAUUUGGGAAUGUGGGCA [428-446](19/19) 54 AGCCAAAACUAGUAUAGAA 557 UUCUAUACUAGUUUUGGCU [2043-2061](19/19) 55 AAAGUAAGAAGCCAGAUGU 558 ACAUCUGGCUUCUUACUUU [1900-1918](19/19) 56 CCAAAAGUAAGAAGCCAGA 559 UCUGGCUUCUUACUUUUGG [1897-1915](19/19) 57 CCCAAAAGUAAGAAGCCAG 560 CUGGCUUCUUACUUUUGGG [1896-1914](19/19) 58 UACGUGAUGAAGAUGGAAA 561 UUUCCAUCUUCAUCACGUA [1816-1834](19/19) 59 GCUCAAAGAAAAAGGAGAA 562 UUCUCCUUUUUCUUUGAGC [1730-1748](19/19) 60 AACUAGAGCAAGAUUUAGA 563 UCUAAAUCUUGCUCUAGUU [1684-1702](19/19) 61 AAGUGGCUGCUCAGAAUUG 564 CAAUUCUGAGCAGCCACUU [1636-1654](19/19) 62 GUAGGAGGGGUAAGAAUAA 565 UUAUUCUUACCCCUCCUAC [1618-1636](19/19) 63 UCAACUUGCAUUAAUUCGG 566 CCGAAUUAAUGCAAGUUGA [1592-1610](19/19) 64 AAAGACAAGAACAACUCCA 567 UGGAGUUGUUCUUGUCUUU [283-301](19/19) 65 AGAAAGAAUUGCCUGUAAG 568 CUUACAGGCAAUUCUUUCU [1393-1411](19/19) 66 ACACACCAGAGAAAGAAUU 569 AAUUCUUUCUCUGGUGUGU [1384-1402](19/19) 67 AACACCAGUACAUUCUUCU 570 AGAAGAAUGUACUGGUGUU [1295-1313](19/19) 68 AAACACCAGUACAUUCUUC 571 GAAGAAUGUACUGGUGUUU [1294-1312](19/19) 69 ACAGAAUGGUCCUAAAACA 572 UGUUUUAGGACCAUUCUGU [1280-1298](19/19) 70 AAACCACCCUGAAAGCACA 573 UGUGCUUUCAGGGUGGUUU [1103-1121](19/19) 71 AACCAAAACCACCCUGAAA 574 UUUCAGGGUGGUUUUGGUU [1098-1116](19/19) 72 UCUCUGAACUUCUAAAUGG 575 CCAUUUAGAAGUUCAGAGA [1039-1057](19/19) 73 UUCACUCUCUGAACUUCUA 576 UAGAAGUUCAGAGAGUGAA [1034-1052](19/19) 74 GUAGUCCACAUUUUCUUAA 577 UUAAGAAAAUGUGGACUAC [838-856](19/19) 75 UUGAAAAUGACAAGCUGGU 578 ACCAGCUUGUCAUUUUCAA [718-736](19/19) 76 CAGUCUUCAUUGCUACUAA 579 UUAGUAGCAAUGAAGACUG [574-592](19/19) 77 AUGACAAUGAGGUUUCUUC 580 GAAGAAACCUCAUUGUCAU [505-523](19/19) 78 AGAUGACAAUGAGGUUUCU 581 AGAAACCUCAUUGUCAUCU [503-521](19/19) 79 GUAGAUGACAAUGAGGUUU 582 AAACCUCAUUGUCAUCUAC [501-519](19/19) 80 UGCCAACUACUCCCAGGUU 583 AACCUGGGAGUAGUUGGCA [410-428](19/19) 81 AAGCCAGAUGUUAAGAAAA 584 UUUUCUUAACAUCUGGCUU [1908-1926](19/19) 82 ACCUUAUUCUCCUAGUGAA 585 UUCACUAGGAGAAUAAGGU [1835-1853](19/19) 83 ACCUACUGAAAAAACAACU 586 AGUUGUUUUUUCAGUAGGU [1765-1783](19/19) 84 ACAAAAGCCUUCACCUACU 587 AGUAGGUGAAGGCUUUUGU [1753-1771](19/19) 85 UGCUCAAAGAAAAAGGAGA 588 UCUCCUUUUUCUUUGAGCA [1729-1747](19/19) 86 CAACUUGCAUUAAUUCGGG 589 CCCGAAUUAAUGCAAGUUG [1593-1611](19/19) 87 GGAAAGACAAGAACAACUC 590 GAGUUGUUCUUGUCUUUCC [281-299](19/19) 88 UUGAAAAGGAAAGACAAGA 591 UCUUGUCUUUCCUUUUCAA [274-292](19/19) 89 CAAAAGACAAACAUUCAAG 592 CUUGAAUGUUUGUCUUUUG [1438-1456](19/19) 90 UCACAAAAGACAAACAUUC 593 GAAUGUUUGUCUUUUGUGA [1435-1453](19/19) 91 ACCCCAUUCACAAAAGACA 594 UGUCUUUUGUGAAUGGGGU [1428-1446](19/19) 92 AGAAUUGCCUGUAAGUCCU 595 AGGACUUACAGGCAAUUCU [1397-1415](19/19) 93 AGCGACGGAAAGAGUAUGA 596 UCAUACUCUUUCCGUCGCU [235-253](19/19) 94 AAGCACAGCAGAAUUCAAU 597 AUUGAAUUCUGCUGUGCUU [1115-1133](19/19) 95 CUCUGAACUUCUAAAUGGG 598 CCCAUUUAGAAGUUCAGAG [1040-1058](19/19) 96 UUGACAGUGAACUCAUUAA 599 UUAAUGAGUUCACUGUCAA [909-927](19/19) 97 UAAGUCGAGAAGUAUUUGA 600 UCAAAUACUUCUCGACUUA [208-226](19/19) 98 UGGAGUAAGUCGAGAAGUA 601 UACUUCUCGACUUACUCCA [203-221](19/19) 99 UACUCAUCUAUACCCUCAA 602 UUGAGGGUAUAGAUGAGUA [798-816](19/19) 100 UCCAAGUCCAGAAGCCAAA 603 UUUGGCUUCUGGACUUGGA [752-770](19/19) 101 CUGUUGAUUUAGACGGUAU 604 AUACCGUCUAAAUCAACAG [634-652](19/19) 102 UUGCUACUAAUCAGGCUCA 605 UGAGCCUGAUUAGUAGCAA [583-601](19/19) 103 GCCCAGUCUUCAUUGCUAC 606 GUAGCAAUGAAGACUGGGC [571-589](19/19) 104 CAUUCCCGUUUGUAGAUGA 607 UCAUCUACAAACGGGAAUG [490-508](19/19) 105 AUGCUUUGUACUUUGAUGA 608 UCAUCAAAGUACAAAGCAU [448-466](19/19) 106 CAGAUGCUUUGUACUUUGA 609 UCAAAGUACAAAGCAUCUG [445-463](19/19) 107 AAGAGCUGGUACUAAUAAA 610 UUUAUUAGUACCAGCUCUU [2370-2388](19/19) 108 GGACAAAAAAUGGCAUUUU 611 AAAAUGCCAUUUUUUGUCC [2310-2328](19/19) 109 GUGAAAUGCUCAUACUUUA 612 UAAAGUAUGAGCAUUUCAC [2001-2019](19/19) 110 AACCUUAUUCUCCUAGUGA 613 UCACUAGGAGAAUAAGGUU [1834-1852](19/19) 111 AAAACCUUAUUCUCCUAGU 614 ACUAGGAGAAUAAGGUUUU [1832-1850](19/19) 112 AAACAACUCAGCACCUUAU 615 AUAAGGUGCUGAGUUGUUU [1776-1794](19/19) 113 CACCUACUGAAAAAACAAC 616 GUUGUUUUUUCAGUAGGUG [1764-1782](19/19) 114 UUCACCUACUGAAAAAACA 617 UGUUUUUUCAGUAGGUGAA [1762-1780](19/19) 115 ACAAGAACAACUCCAAAAG 618 CUUUUGGAGUUGUUCUUGU [287-305](19/19) 116 UUGCAUUAAUUCGGGAUAU 619 AUAUCCCGAAUUAAUGCAA [1597-1615](19/19) 117 AACUUGCAUUAAUUCGGGA 620 UCCCGAAUUAAUGCAAGUU [1594-1612](19/19) 118 CACAAAAGACAAACAUUCA 621 UGAAUGUUUGUCUUUUGUG [1436-1454](19/19) 119 CCCAUUCACAAAAGACAAA 622 UUUGUCUUUUGUGAAUGGG [1430-1448](19/19) 120 GAACACACCAGAGAAAGAA 623 UUCUUUCUCUGGUGUGUUC [1382-1400](19/19) 121 ACGGUCCACAGCUCAUCAU 624 AUGAUGAGCUGUGGACCGU [97-115](19/19) 122 UGAGAACACACCAGAGAAA 625 UUUCUCUGGUGUGUUCUCA [1379-1397](19/19) 123 GUGAUUCUGAAGUGGAAGA 626 UCUUCCACUUCAGAAUCAC [1234-1252](19/19) 124 ACAGCAGAAUUCAAUGAUU 627 AAUCAUUGAAUUCUGCUGU [1119-1137](19/19) 125 GCACAGCAGAAUUCAAUGA 628 UCAUUGAAUUCUGCUGUGC [1117-1135](19/19) 126 CUCAUUAAAUUCAGAUGCC 629 GGCAUCUGAAUUUAAUGAG [920-938](19/19) 127 ACUCAUUAAAUUCAGAUGC 630 GCAUCUGAAUUUAAUGAGU [919-937](19/19) 128 AACCAGUUGACAGUGAACU 631 AGUUCACUGUCAACUGGUU [903-921](19/19) 129 UCCUUCAGCAGCAUCCUCU 632 AGAGGAUGCUGCUGAAGGA [870-888](19/19) 130 GAGGAUUCCUUCAGCAGCA 633 UGCUGCUGAAGGAAUCCUC [864-882](19/19) 131 AGUAAGUCGAGAAGUAUUU 634 AAAUACUUCUCGACUUACU [206-224](19/19) 132 GAGUAAGUCGAGAAGUAUU 635 AAUACUUCUCGACUUACUC [205-223](19/19) 133 ACUCAUCUAUACCCUCAAU 636 AUUGAGGGUAUAGAUGAGU [799-817](19/19) 134 UUACUCAUCUAUACCCUCA 637 UGAGGGUAUAGAUGAGUAA [797-815](19/19) 135 UGAGUUACAGUGUCUUAAU 638 AUUAAGACACUGUAACUCA [698-716](19/19) 136 UUUGGAGGCAAGAUAUAGA 639 UCUAUAUCUUGCCUCCAAA [181-199](19/19) 137 AGGACAUUGAGCAAGUUUG 640 CAAACUUGCUCAAUGUCCU [658-676](19/19) 138 UAGACGGUAUGCAACAGGA 641 UCCUGUUGCAUACCGUCUA [643-661](19/19) 139 UUGAUUUAGACGGUAUGCA 642 UGCAUACCGUCUAAAUCAA [637-655](19/19) 140 UUCAUUGCUACUAAUCAGG 643 CCUGAUUAGUAGCAAUGAA [579-597](19/19) 141 AGUCUUCAUUGCUACUAAU 644 AUUAGUAGCAAUGAAGACU [575-593](19/19) 142 CGGCUACGUUUCAGUCACU 645 AGUGACUGAAACGUAGCCG [523-541](19/19) 143 AGGACAUGGAUUUGAUUGA 646 UCAAUCAAAUCCAUGUCCU [157-175](19/19) 144 GCCCACAUUCCCAAAUCAG 647 CUGAUUUGGGAAUGUGGGC [429-447](19/19) 145 AGCAGGACAUGGAUUUGAU 648 AUCAAAUCCAUGUCCUGCU [154-172](19/19) 146 AGCUGGUACUAAUAAAGGA 649 UCCUUUAUUAGUACCAGCU [2373-2391](19/19) 147 GAGCUGGUACUAAUAAAGG 650 CCUUUAUUAGUACCAGCUC [2372-2390](19/19) 148 AGAGCUGGUACUAAUAAAG 651 CUUUAUUAGUACCAGCUCU [2371-2389](19/19) 149 CUAACCACUAUGUACUUUU 652 AAAAGUACAUAGUGGUUAG [2276-2294](19/19) 150 ACAUCUGGCUAAAAAGAAA 653 UUUCUUUUUAGCCAGAUGU [2246-2264](19/19) 151 CUGUUCUUAUGUCAUUUGU 654 ACAAAUGACAUAAGAACAG [2199-2217](19/19) 152 UGGGCUAGUUUCUGUGUAA 655 UUACACAGAAACUAGCCCA [2152-2170](19/19) 153 GUAAACAAUUUCUUAGGAC 656 GUCCUAAGAAAUUGUUUAC [2127-2145](19/19) 154 CUGUAAACAAUUUCUUAGG 657 CCUAAGAAAUUGUUUACAG [2125-2143](19/19) 155 GUCAGUAUGUUGAAUCAGU 658 ACUGAUUCAACAUACUGAC [2093-2111](19/19) 156 GGCAAUGUUUUCCUUGUUC 659 GAACAAGGAAAACAUUGCC [1878-1896](19/19) 157 GCACCUUAUAUCUCGAAGU 660 ACUUCGAGAUAUAAGGUGC [1786-1804](19/19) 158 GAAAAAACAACUCAGCACC 661 GGUGCUGAGUUGUUUUUUC [1772-1790](19/19) 159 AGAGCAAGAUUUAGAUCAU 662 AUGAUCUAAAUCUUGCUCU [1688-1706](19/19) 160 UAGAGCAAGAUUUAGAUCA 663 UGAUCUAAAUCUUGCUCUA [1687-1705](19/19) 161 UAGAACUAGAGCAAGAUUU 664 AAAUCUUGCUCUAGUUCUA [1681-1699](19/19) 162 ACGUAGGAGGGGUAAGAAU 665 AUUCUUACCCCUCCUACGU [1616-1634](19/19) 163 UCCCUGUUGUUGACUUCAA 666 UUGAAGUCAACAACAGGGA [1540-1558](19/19) 164 UCCCUGUAGAAAAAAUCAU 667 AUGAUUUUUUCUACAGGGA [1516-1534](19/19) 165 UCACAAGAGAUGAACUUAG 668 CUAAGUUCAUCUCUUGUGA [1474-1492](19/19) 166 AGAGAAAGAAUUGCCUGUA 669 UACAGGCAAUUCUUUCUCU [1391-1409](19/19) 167 ACCAGAGAAAGAAUUGCCU 670 AGGCAAUUCUUUCUCUGGU [1388-1406](19/19) 168 CACCAGAGAAAGAAUUGCC 671 GGCAAUUCUUUCUCUGGUG [1387-1405](19/19) 169 AGUAUGAGCUGGAAAAACA 672 UGUUUUUCCAGCUCAUACU [247-265](19/19) 170 GUACAUUCUUCUGGGGAUA 673 UAUCCCCAGAAGAAUGUAC [1302-1320](19/19) 171 CAAACAGAAUGGUCCUAAA 674 UUUAGGACCAUUCUGUUUG [1277-1295](19/19) 172 CUGGAAGUGUCAAACAGAA 675 UUCUGUUUGACACUUCCAG [1267-1285](19/19) 173 GGAAAGAGUAUGAGCUGGA 676 UCCAGCUCAUACUCUUUCC [241-259](19/19) 174 CAGUGAUUCUGAAGUGGAA 677 UUCCACUUCAGAAUCACUG [1232-1250](19/19) 175 GACGGAAAGAGUAUGAGCU 678 AGCUCAUACUCUUUCCGUC [238-256](19/19) 176 CACAGCAGAAUUCAAUGAU 679 AUCAUUGAAUUCUGCUGUG [1118-1136](19/19) 177 GCCACAGUCAACACAGAUU 680 AAUCUGUGUUGACUGUGGC [936-954](19/19) 178 UAAAUUCAGAUGCCACAGU 681 ACUGUGGCAUCUGAAUUUA [925-943](19/19) 179 CCAGUUGACAGUGAACUCA 682 UGAGUUCACUGUCAACUGG [905-923](19/19) 180 GUCGAGAAGUAUUUGACUU 683 AAGUCAAAUACUUCUCGAC [211-229](19/19) 181 CUGUAGUCCACAUUUUCUU 684 AAGAAAAUGUGGACUACAG [836-854](19/19) 182 GGAGUAAGUCGAGAAGUAU 685 AUACUUCUCGACUUACUCC [204-222](19/19) 183 GACAGAAGUUGACAAUUAU 686 AUAAUUGUCAACUUCUGUC [773-791](19/19) 184 GAGACUACCAUGGUUCCAA 687 UUGGAACCAUGGUAGUCUC [738-756](19/19) 185 CUAUUAUCCAUUCCUGAGU 688 ACUCAGGAAUGGAUAAUAG [684-702](19/19) 186 GACGGUAUGCAACAGGACA 689 UGUCCUGUUGCAUACCGUC [645-663](19/19) 187 CGAGAGCCCAGUCUUCAUU 690 AAUGAAGACUGGGCUCUCG [566-584](19/19) 188 UUGAUUGACAUACUUUGGA 691 UCCAAAGUAUGUCAAUCAA [168-186](19/19) 189 GUCACUUGUUCCUGAUAUU 692 AAUAUCAGGAACAAGUGAC [536-554](19/19) 190 UCGGCUACGUUUCAGUCAC 693 GUGACUGAAACGUAGCCGA [522-540](19/19) 191 GAGGUUUCUUCGGCUACGU 694 ACGUAGCCGAAGAAACCUC [513-531](19/19) 192 GACAUUCCCGUUUGUAGAU 695 AUCUACAAACGGGAAUGUC [488-506](19/19) 193 GACAUGGAUUUGAUUGACA 696 UGUCAAUCAAAUCCAUGUC [159-177](19/19) 194 UCAGAUGCUUUGUACUUUG 697 CAAAGUACAAAGCAUCUGA [444-462](19/19) 195 CAGCAGGACAUGGAUUUGA 698 UCAAAUCCAUGUCCUGCUG [153-171](19/19) 196 AUGGACAAAAAAUGGCAUU 699 AAUGCCAUUUUUUGUCCAU [2308-2326](19/19) 197 GUAUGGACAAAAAAUGGCA 700 UGCCAUUUUUUGUCCAUAC [2306-2324](19/19) 198 CAUCUGGCUAAAAAGAAAU 701 AUUUCUUUUUAGCCAGAUG [2247-2265](19/19) 199 GACAUCUGGCUAAAAAGAA 702 UUCUUUUUAGCCAGAUGUC [2245-2263](19/19) 200 GAGCUAGUUUUUUUGUACU 703 AGUACAAAAAAACUAGCUC [1956-1974](19/19) 201 GAAGAUGGAAAACCUUAUU 704 AAUAAGGUUUUCCAUCUUC [1824-1842](19/19) 202 GCUACGUGAUGAAGAUGGA 705 UCCAUCUUCAUCACGUAGC [1814-1832](19/19) 203 UGCUACGUGAUGAAGAUGG 706 CCAUCUUCAUCACGUAGCA [1813-1831](19/19) 204 GAGCAAGAUUUAGAUCAUU 707 AAUGAUCUAAAUCUUGCUC [1689-1707](19/19) 205 ACUAGAGCAAGAUUUAGAU 708 AUCUAAAUCUUGCUCUAGU [1685-1703](19/19) 206 UAGUAGAACUAGAGCAAGA 709 UCUUGCUCUAGUUCUACUA [1678-1696](19/19) 207 AUAUCCCAUUCCCUGUAGA 710 UCUACAGGGAAUGGGAUAU [1507-1525](19/19) 208 CCAUAUCCCAUUCCCUGUA 711 UACAGGGAAUGGGAUAUGG [1505-1523](19/19) 209 CCCCAUUCACAAAAGACAA 712 UUGUCUUUUGUGAAUGGGG [1429-1447](19/19) 210 ACACCAGAGAAAGAAUUGC 713 GCAAUUCUUUCUCUGGUGU [1386-1404](19/19) 211 UCAAACAGAAUGGUCCUAA 714 UUAGGACCAUUCUGUUUGA [1276-1294](19/19) 212 AAGAGUAUGAGCUGGAAAA 715 UUUUCCAGCUCAUACUCUU [244-262](19/19) 213 UGCCCCUGGAAGUGUCAAA 716 UUUGACACUUCCAGGGGCA [1262-1280](19/19) 214 GCGACGGAAAGAGUAUGAG 717 CUCAUACUCUUUCCGUCGC [236-254](19/19) 215 CUGAACUUCUAAAUGGGCC 718 GGCCCAUUUAGAAGUUCAG [1042-1060](19/19) 216 CCACAGUCAACACAGAUUU 719 AAAUCUGUGUUGACUGUGG [937-955](19/19) 217 AUGCUUUUGAGGAUUCCUU 720 AAGGAAUCCUCAAAAGCAU [856-874](19/19) 218 CUCAAUGGAAAAAGAAGUA 721 UACUUCUUUUUCCAUUGAG [812-830](19/19) 219 CUCAUCUAUACCCUCAAUG 722 CAUUGAGGGUAUAGAUGAG [800-818](19/19) 220 ACAUUGAGCAAGUUUGGGA 723 UCCCAAACUUGCUCAAUGU [661-679](19/19) 221 AACAGGACAUUGAGCAAGU 724 ACUUGCUCAAUGUCCUGUU [655-673](19/19) 222 UUAGACGGUAUGCAACAGG 725 CCUGUUGCAUACCGUCUAA [642-660](19/19) 223 ACCUGAAACUUCUGUUGCU 726 AGCAACAGAAGUUUCAGGU [605-623](19/19) 224 CCAGUCUUCAUUGCUACUA 727 UAGUAGCAAUGAAGACUGG [573-591](19/19) 225 AGAGCCCAGUCUUCAUUGC 728 GCAAUGAAGACUGGGCUCU [568-586](19/19) 226 AGUCACUUGUUCCUGAUAU 729 AUAUCAGGAACAAGUGACU [535-553](19/19) 227 UGGAUUUGAUUGACAUACU 730 AGUAUGUCAAUCAAAUCCA [163-181](19/19) 228 CAAUGAGGUUUCUUCGGCU 731 AGCCGAAGAAACCUCAUUG [509-527](19/19) 229 ACAAUGAGGUUUCUUCGGC 732 GCCGAAGAAACCUCAUUGU [508-526](19/19) 230 AGACAUUCCCGUUUGUAGA 733 UCUACAAACGGGAAUGUCU [487-505](19/19) 231 GCUUUGUACUUUGAUGACU 734 AGUCAUCAAAGUACAAAGC [450-468](19/19) 232 CCCACAUUCCCAAAUCAGA 735 UCUGAUUUGGGAAUGUGGG [430-448](19/19) 233 ACAACUAGAUGAAGAGACA 736 UGUCUCUUCAUCUAGUUGU [335-353](19/19) 234 UACAACUAGAUGAAGAGAC 737 GUCUCUUCAUCUAGUUGUA [334-352](19/19) 235 UUUAAGAGCUGGUACUAAU 738 AUUAGUACCAGCUCUUAAA [2367-2385](19/19) 236 UGGACAAAAAAUGGCAUUU 739 AAAUGCCAUUUUUUGUCCA [2309-2327](19/19) 237 GGGCUAGUUUCUGUGUAAG 740 CUUACACAGAAACUAGCCC [2153-2171](19/19) 238 UUGGGCUAGUUUCUGUGUA 741 UACACAGAAACUAGCCCAA [2151-2169](19/19) 239 GUGUCAGUAUGUUGAAUCA 742 UGAUUCAACAUACUGACAC [2091-2109](19/19) 240 GAUGUGAAAUGCUCAUACU 743 AGUAUGAGCAUUUCACAUC [1998-2016](19/19) 241 CUCCUACUGUGAUGUGAAA 744 UUUCACAUCACAGUAGGAG [1988-2006](19/19) 242 AGAUUUAGGAGGAUUUGAC 745 GUCAAAUCCUCCUAAAUCU [1930-1948](19/19) 243 GCAAUGUUUUCCUUGUUCC 746 GGAACAAGGAAAACAUUGC [1879-1897](19/19) 244 AACAAGAGAUGGCAAUGUU 747 AACAUUGCCAUCUCUUGUU [1868-1886](19/19) 245 GCAGCAAACAAGAGAUGGC 748 GCCAUCUCUUGUUUGCUGC [1862-1880](19/19) 246 CUCCUAGUGAAUACUCCCU 749 AGGGAGUAUUCACUAGGAG [1843-1861](19/19) 247 CAGCACCUUAUAUCUCGAA 750 UUCGAGAUAUAAGGUGCUG [1784-1802](19/19) 248 UCACCUACUGAAAAAACAA 751 UUGUUUUUUCAGUAGGUGA [1763-1781](19/19) 249 GCAAGAUUUAGAUCAUUUG 752 CAAAUGAUCUAAAUCUUGC [1691-1709](19/19) 250 CAGAAAAAGAAAACUGGAA 753 UUCCAGUUUUCUUUUUCUG [1655-1673](19/19) 251 AUACGUAGGAGGGGUAAGA 754 UCUUACCCCUCCUACGUAU [1614-1632](19/19) 252 UGCAUUAAUUCGGGAUAUA 755 UAUAUCCCGAAUUAAUGCA [1598-1616](19/19) 253 AAGCUCUCCAUAUCCCAUU 756 AAUGGGAUAUGGAGAGCUU [1498-1516](19/19) 254 CAUUCACAAAAGACAAACA 757 UGUUUGUCUUUUGUGAAUG [1432-1450](19/19) 255 GAGUAUGAGCUGGAAAAAC 758 GUUUUUCCAGCUCAUACUC [246-264](19/19) 256 CAGAAUGGUCCUAAAACAC 759 GUGUUUUAGGACCAUUCUG [1281-1299](19/19) 257 GUGUCAAACAGAAUGGUCC 760 GGACCAUUCUGUUUGACAC [1273-1291](19/19) 258 GAAGUGUCAAACAGAAUGG 761 CCAUUCUGUUUGACACUUC [1270-1288](19/19) 259 CUGAAGUGGAAGAGCUAGA 762 UCUAGCUCUUCCACUUCAG [1240-1258](19/19) 260 CCAGAACACUCAGUGGAAU 763 AUUCCACUGAGUGUUCUGG [1182-1200](19/19) 261 UUCUGACUCCGGCAUUUCA 764 UGAAAUGCCGGAGUCAGAA [1136-1154](19/19) 262 AUGAUUCUGACUCCGGCAU 765 AUGCCGGAGUCAGAAUCAU [1132-1150](19/19) 263 GGCCCAUUGAUGUUUCUGA 766 UCAGAAACAUCAAUGGGCC [1057-1075](19/19) 264 CACAGAUUUUGGUGAUGAA 767 UUCAUCACCAAAAUCUGUG [947-965](19/19) 265 CAGUUGACAGUGAACUCAU 768 AUGAGUUCACUGUCAACUG [906-924](19/19) 266 UUAAUGCUUUUGAGGAUUC 769 GAAUCCUCAAAAGCAUUAA [853-871](19/19) 267 UCUUAAUGCUUUUGAGGAU 770 AUCCUCAAAAGCAUUAAGA [851-869](19/19) 268 AGUCCACAUUUUCUUAAUG 771 CAUUAAGAAAAUGUGGACU [840-858](19/19) 269 UGACAGAAGUUGACAAUUA 772 UAAUUGUCAACUUCUGUCA [772-790](19/19) 270 CCAAACUGACAGAAGUUGA 773 UCAACUUCUGUCAGUUUGG [766-784](19/19) 271 GUUGAGACUACCAUGGUUC 774 GAACCAUGGUAGUCUCAAC [735-753](19/19) 272 ACAAGCUGGUUGAGACUAC 775 GUAGUCUCAACCAGCUUGU [727-745](19/19) 273 AUACUUUGGAGGCAAGAUA 776 UAUCUUGCCUCCAAAGUAU [177-195](19/19) 274 GACAUACUUUGGAGGCAAG 777 CUUGCCUCCAAAGUAUGUC [174-192](19/19) 275 GUCACCUGAAACUUCUGUU 778 AACAGAAGUUUCAGGUGAC [602-620](19/19) 276 UCGAGAGCCCAGUCUUCAU 779 AUGAAGACUGGGCUCUCGA [565-583](19/19) 277 GUACUUUGAUGACUGCAUG 780 CAUGCAGUCAUCAAAGUAC [455-473](19/19) 278 UGUACUUUGAUGACUGCAU 781 AUGCAGUCAUCAAAGUACA [454-472](19/19) 279 GGACAUGGAUUUGAUUGAC 782 GUCAAUCAAAUCCAUGUCC [158-176](19/19) 280 GAUGCUUUGUACUUUGAUG 783 CAUCAAAGUACAAAGCAUC [447-465](19/19) 281 AUCAGAUGCUUUGUACUUU 784 AAAGUACAAAGCAUCUGAU [443-461](19/19) 282 UUCCCAAAUCAGAUGCUUU 785 AAAGCAUCUGAUUUGGGAA [436-454](19/19) 283 CAUUCCCAAAUCAGAUGCU 786 AGCAUCUGAUUUGGGAAUG [434-452](19/19) 284 CCACAUUCCCAAAUCAGAU 787 AUCUGAUUUGGGAAUGUGG [431-449](19/19) 285 AGUUACAACUAGAUGAAGA 788 UCUUCAUCUAGUUGUAACU [331-349](19/19) 286 UAUGGACAAAAAAUGGCAU 789 AUGCCAUUUUUUGUCCAUA [2307-2325](19/19) 287 ACUGUAUGGACAAAAAAUG 790 CAUUUUUUGUCCAUACAGU [2303-2321](19/19) 288 UAUGACAUCUGGCUAAAAA 791 UUUUUAGCCAGAUGUCAUA [2242-2260](19/19) 289 ACUGUUCUUAUGUCAUUUG 792 CAAAUGACAUAAGAACAGU [2198-2216](19/19) 290 UAGUUUCUGUGUAAGUGUA 793 UACACUUACACAGAAACUA [2157-2175](19/19) 291 GGCUAGUUUCUGUGUAAGU 794 ACUUACACAGAAACUAGCC [2154-2172](19/19) 292 CAAUUUCUUAGGACACCAU 795 AUGGUGUCCUAAGAAAUUG [2132-2150](19/19) 293 GUAUGUUGAAUCAGUAGUU 796 AACUACUGAUUCAACAUAC [2097-2115](19/19) 294 UCAGUAUGUUGAAUCAGUA 797 UACUGAUUCAACAUACUGA [2094-2112](19/19) 295 UGAUGUGAAAUGCUCAUAC 798 GUAUGAGCAUUUCACAUCA [1997-2015](19/19) 296 AAAGCUCCUACUGUGAUGU 799 ACAUCACAGUAGGAGCUUU [1984-2002](19/19) 297 UACUAAAAGCUCCUACUGU 800 ACAGUAGGAGCUUUUAGUA [1979-1997](19/19) 298 GGAUUUGACCUUUUCUGAG 801 CUCAGAAAAGGUCAAAUCC [1940-1958](19/19) 299 AGAGAUGGCAAUGUUUUCC 802 GGAAAACAUUGCCAUCUCU [1872-1890](19/19) 300 AAACAAGAGAUGGCAAUGU 803 ACAUUGCCAUCUCUUGUUU [1867-1885](19/19) 301 CAGCAAACAAGAGAUGGCA 804 UGCCAUCUCUUGUUUGCUG [1863-1881](19/19) 302 CUAGUGAAUACUCCCUGCA 805 UGCAGGGAGUAUUCACUAG [1846-1864](19/19) 303 AAGAUGGAAAACCUUAUUC 806 GAAUAAGGUUUUCCAUCUU [1825-1843](19/19) 304 CAGCAUGCUACGUGAUGAA 807 UUCAUCACGUAGCAUGCUG [1808-1826](19/19) 305 CACCUUAUAUCUCGAAGUU 808 AACUUCGAGAUAUAAGGUG [1787-1805](19/19) 306 UCAGCACCUUAUAUCUCGA 809 UCGAGAUAUAAGGUGCUGA [1783-1801](19/19) 307 AUAUAGUAGAACUAGAGCA 810 UGCUCUAGUUCUACUAUAU [1675-1693](19/19) 308 UUGCAGAAAAAGAAAACUG 811 CAGUUUUCUUUUUCUGCAA [1652-1670](19/19) 309 ACGAAAUGAUGUCCAAAGA 812 UCUUUGGACAUCAUUUCGU [1558-1576](19/19) 310 ACCUCCCUGUUGUUGACUU 813 AAGUCAACAACAGGGAGGU [1537-1555](19/19) 311 CCCUGUAGAAAAAAUCAUU 814 AAUGAUUUUUUCUACAGGG [1517-1535](19/19) 312 GAGAUGAACUUAGGGCAAA 815 UUUGCCCUAAGUUCAUCUC [1480-1498](19/19) 313 CACAAGAGAUGAACUUAGG 816 CCUAAGUUCAUCUCUUGUG [1475-1493](19/19) 314 AACCCCAUUCACAAAAGAC 817 GUCUUUUGUGAAUGGGGUU [1427-1445](19/19) 315 AGAACACACCAGAGAAAGA 818 UCUUUCUCUGGUGUGUUCU [1381-1399](19/19) 316 GUCAAACAGAAUGGUCCUA 819 UAGGACCAUUCUGUUUGAC [1275-1293](19/19) 317 GGAAGUGUCAAACAGAAUG 820 CAUUCUGUUUGACACUUCC [1269-1287](19/19) 318 UGGAAGUGUCAAACAGAAU 821 AUUCUGUUUGACACUUCCA [1268-1286](19/19) 319 CCCUGGAAGUGUCAAACAG 822 CUGUUUGACACUUCCAGGG [1265-1283](19/19) 320 GUGGAAGAGCUAGAUAGUG 823 CACUAUCUAGCUCUUCCAC [1245-1263](19/19) 321 ACCAGAACACUCAGUGGAA 824 UUCCACUGAGUGUUCUGGU [1181-1199](19/19) 322 CAGCGACGGAAAGAGUAUG 825 CAUACUCUUUCCGUCGCUG [234-252](19/19) 323 UGAUCUAUCACUUUGCAAA 826 UUUGCAAAGUGAUAGAUCA [1073-1091](19/19) 324 GCCCAUUGAUGUUUCUGAU 827 AUCAGAAACAUCAAUGGGC [1058-1076](19/19) 325 AGUCAACACAGAUUUUGGU 828 ACCAAAAUCUGUGUUGACU [941-959](19/19) 326 AGUUGACAGUGAACUCAUU 829 AAUGAGUUCACUGUCAACU [907-925](19/19) 327 ACUGUAGUCCACAUUUUCU 830 AGAAAAUGUGGACUACAGU [835-853](19/19) 328 CAAUGGAAAAAGAAGUAGG 831 CCUACUUCUUUUUCCAUUG [814-832](19/19) 329 CAAACUGACAGAAGUUGAC 832 GUCAACUUCUGUCAGUUUG [767-785](19/19) 330 AGACUACCAUGGUUCCAAG 833 CUUGGAACCAUGGUAGUCU [739-757](19/19) 331 ACAGGACAUUGAGCAAGUU 834 AACUUGCUCAAUGUCCUGU [656-674](19/19) 332 UGACAUACUUUGGAGGCAA 835 UUGCCUCCAAAGUAUGUCA [173-191](19/19) 333 ACGUUUCAGUCACUUGUUC 836 GAACAAGUGACUGAAACGU [528-546](19/19) 334 CUACGUUUCAGUCACUUGU 837 ACAAGUGACUGAAACGUAG [526-544](19/19) 335 GGCUACGUUUCAGUCACUU 838 AAGUGACUGAAACGUAGCC [524-542](19/19) 336 GGAUUUGAUUGACAUACUU 839 AAGUAUGUCAAUCAAAUCC [164-182](19/19) 337 UCUUCGGCUACGUUUCAGU 840 ACUGAAACGUAGCCGAAGA [519-537](19/19) 338 CAGUCAGAAACCAGUGGAU 841 AUCCACUGGUUUCUGACUG [390-408](19/19) 339 UGAAGAGACAGGUGAAUUU 842 AAAUUCACCUGUCUCUUCA [344-362](19/19) 340 UAGAUGAAGAGACAGGUGA 843 UCACCUGUCUCUUCAUCUA [340-358](19/19) 341 UAAGAGCUGGUACUAAUAA 844 UUAUUAGUACCAGCUCUUA [2369-2387](19/19) 342 CUGUAUGGACAAAAAAUGG 845 CCAUUUUUUGUCCAUACAG [2304-2322](19/19) 343 UGGAGUGUCAGUAUGUUGA 846 UCAACAUACUGACACUCCA [2087-2105](19/19) 344 UGUGAAAUGCUCAUACUUU 847 AAAGUAUGAGCAUUUCACA [2000-2018](19/19) 345 GUGAUGUGAAAUGCUCAUA 848 UAUGAGCAUUUCACAUCAC [1996-2014](19/19) 346 AGCUCCUACUGUGAUGUGA 849 UCACAUCACAGUAGGAGCU [1986-2004](19/19) 347 UCCUAGUGAAUACUCCCUG 850 CAGGGAGUAUUCACUAGGA [1844-1862](19/19) 348 UUCUCCUAGUGAAUACUCC 851 GGAGUAUUCACUAGGAGAA [1841-1859](19/19) 349 CAUGCUACGUGAUGAAGAU 852 AUCUUCAUCACGUAGCAUG [1811-1829](19/19) 350 GCAUGCUACGUGAUGAAGA 853 UCUUCAUCACGUAGCAUGC [1810-1828](19/19) 351 AGCACCUUAUAUCUCGAAG 854 CUUCGAGAUAUAAGGUGCU [1785-1803](19/19) 352 AGAAAAAGGAGAAAAUGAC 855 GUCAUUUUCUCCUUUUUCU [1736-1754](19/19) 353 UGCAGAAAAAGAAAACUGG 856 CCAGUUUUCUUUUUCUGCA [1653-1671](19/19) 354 UCAUUAACCUCCCUGUUGU 857 ACAACAGGGAGGUUAAUGA [1531-1549](19/19) 355 AAGUGUCAAACAGAAUGGU 858 ACCAUUCUGUUUGACACUU [1271-1289](19/19) 356 GAAAGAGUAUGAGCUGGAA 859 UUCCAGCUCAUACUCUUUC [242-260](19/19) 357 AAAGCACAGCAGAAUUCAA 860 UUGAAUUCUGCUGUGCUUU [1114-1132](19/19) 358 AUGUUUCUGAUCUAUCACU 861 AGUGAUAGAUCAGAAACAU [1066-1084](19/19) 359 UGGGCCCAUUGAUGUUUCU 862 AGAAACAUCAAUGGGCCCA [1055-1073](19/19) 360 CAGUCAACACAGAUUUUGG 863 CCAAAAUCUGUGUUGACUG [940-958](19/19) 361 ACAGUCAACACAGAUUUUG 864 CAAAAUCUGUGUUGACUGU [939-957](19/19) 362 CACAGUCAACACAGAUUUU 865 AAAAUCUGUGUUGACUGUG [938-956](19/19) 363 ACAGAAGACCCCAACCAGU 866 ACUGGUUGGGGUCUUCUGU [891-909](19/19) 364 AGGAUUCCUUCAGCAGCAU 867 AUGCUGCUGAAGGAAUCCU [865-883](19/19) 365 AAUGGAAAAAGAAGUAGGU 868 ACCUACUUCUUUUUCCAUU [815-833](19/19) 366 AUGACAAGCUGGUUGAGAC 869 GUCUCAACCAGCUUGUCAU [724-742](19/19) 367 UUGAGCAAGUUUGGGAGGA 870 UCCUCCCAAACUUGCUCAA [664-682](19/19) 368 CUGAAACUUCUGUUGCUCA 871 UGAGCAACAGAAGUUUCAG [607-625](19/19) 369 UCACCUGAAACUUCUGUUG 872 CAACAGAAGUUUCAGGUGA [603-621](19/19) 370 UUGACAUACUUUGGAGGCA 873 UGCCUCCAAAGUAUGUCAA [172-190](19/19) 371 UGAUUGACAUACUUUGGAG 874 CUCCAAAGUAUGUCAAUCA [169-187](19/19) 372 AGGUUUCUUCGGCUACGUU 875 AACGUAGCCGAAGAAACCU [514-532](19/19) 373 AUGGAUUUGAUUGACAUAC 876 GUAUGUCAAUCAAAUCCAU [162-180](19/19) 374 ACAUGGAUUUGAUUGACAU 877 AUGUCAAUCAAAUCCAUGU [160-178](19/19) 375 UUCUCCCAAUUCAGCCAGC 878 GCUGGCUGAAUUGGGAGAA [361-379](19/19) 376 AAGAGACAGGUGAAUUUCU 879 AGAAAUUCACCUGUCUCUU [346-364](19/19) 377 GAUGAAGAGACAGGUGAAU 880 AUUCACCUGUCUCUUCAUC [342-360](19/19) 378 AACUAGAUGAAGAGACAGG 881 CCUGUCUCUUCAUCUAGUU [337-355](19/19) 379 UUGCAAAACUAACCACUAU 882 AUAGUGGUUAGUUUUGCAA [2268-2286](19/19) 380 UGACAUCUGGCUAAAAAGA 883 UCUUUUUAGCCAGAUGUCA [2244-2262](19/19) 381 GAGUGUCAGUAUGUUGAAU 884 AUUCAACAUACUGACACUC [2089-2107](19/19) 382 AAGCAUUGGAGUGUCAGUA 885 UACUGACACUCCAAUGCUU [2081-2099](19/19) 383 GCUCCUACUGUGAUGUGAA 886 UUCACAUCACAGUAGGAGC [1987-2005](19/19) 384 GACCUUUUCUGAGCUAGUU 887 AACUAGCUCAGAAAAGGUC [1946-1964](19/19) 385 UGACCUUUUCUGAGCUAGU 888 ACUAGCUCAGAAAAGGUCA [1945-1963](19/19) 386 UCAUGAUGGACUUGGAGCU 889 AGCUCCAAGUCCAUCAUGA [112-130](19/19) 387 CCUUAUAUCUCGAAGUUUU 890 AAAACUUCGAGAUAUAAGG [1789-1807](19/19) 388 CUCAGCACCUUAUAUCUCG 891 CGAGAUAUAAGGUGCUGAG [1782-1800](19/19) 389 GACAAGAACAACUCCAAAA 892 UUUUGGAGUUGUUCUUGUC [286-304](19/19) 390 UCGGGAUAUACGUAGGAGG 893 CCUCCUACGUAUAUCCCGA [1607-1625](19/19) 391 UAAUUCGGGAUAUACGUAG 894 CUACGUAUAUCCCGAAUUA [1603-1621](19/19) 392 GAAAUGAUGUCCAAAGAGC 895 GCUCUUUGGACAUCAUUUC [1560-1578](19/19) 393 AACGAAAUGAUGUCCAAAG 896 CUUUGGACAUCAUUUCGUU [1557-1575](19/19) 394 GAAAAAAUCAUUAACCUCC 897 GGAGGUUAAUGAUUUUUUC [1524-1542](19/19) 395 AGCUCUCCAUAUCCCAUUC 898 GAAUGGGAUAUGGAGAGCU [1499-1517](19/19) 396 GCAAAAGCUCUCCAUAUCC 899 GGAUAUGGAGAGCUUUUGC [1494-1512](19/19) 397 AAGAGAUGAACUUAGGGCA 900 UGCCCUAAGUUCAUCUCUU [1478-1496](19/19) 398 CUGGAAAAACAGAAAAAAC 901 GUUUUUUCUGUUUUUCCAG [255-273](19/19) 399 GAGCUGGAAAAACAGAAAA 902 UUUUCUGUUUUUCCAGCUC [252-270](19/19) 400 CCCCUGGAAGUGUCAAACA 903 UGUUUGACACUUCCAGGGG [1264-1282](19/19) 401 AUUCUGAAGUGGAAGAGCU 904 AGCUCUUCCACUUCAGAAU [1237-1255](19/19) 402 CAUCACCAGAACACUCAGU 905 ACUGAGUGUUCUGGUGAUG [1177-1195](19/19) 403 CUGAUCUAUCACUUUGCAA 906 UUGCAAAGUGAUAGAUCAG [1072-1090](19/19) 404 UCUGAUCUAUCACUUUGCA 907 UGCAAAGUGAUAGAUCAGA [1071-1089](19/19) 405 UUCUGAUCUAUCACUUUGC 908 GCAAAGUGAUAGAUCAGAA [1070-1088](19/19) 406 GAGCCCAGUAUCAGCAACA 909 UGUUGCUGAUACUGGGCUC [987-1005](19/19) 407 GAAGUAGGUAACUGUAGUC 910 GACUACAGUUACCUACUUC [825-843](19/19) 408 AGAAGUAGGUAACUGUAGU 911 ACUACAGUUACCUACUUCU [824-842](19/19) 409 AAAAGAAGUAGGUAACUGU 912 ACAGUUACCUACUUCUUUU [821-839](19/19) 410 GCAACAGGACAUUGAGCAA 913 UUGCUCAAUGUCCUGUUGC [653-671](19/19) 411 CACUUGUUCCUGAUAUUCC 914 GGAAUAUCAGGAACAAGUG [538-556](19/19) 412 GCAGGACAUGGAUUUGAUU 915 AAUCAAAUCCAUGUCCUGC [155-173](19/19) 413 GUUUCUGUGUAAGUGUAAA 916 UUUACACUUACACAGAAAC [2159-2177](19/19) 414 GCAUUGGAGUGUCAGUAUG 917 CAUACUGACACUCCAAUGC [2083-2101](19/19) 415 UGUGAUGUGAAAUGCUCAU 918 AUGAGCAUUUCACAUCACA [1995-2013](19/19) 416 ACUAGAUUUAGGAGGAUUU 919 AAAUCCUCCUAAAUCUAGU [1927-1945](19/19) 417 CUCGAAGUUUUCAGCAUGC 920 GCAUGCUGAAAACUUCGAG [1797-1815](19/19) 418 AUCUCGAAGUUUUCAGCAU 921 AUGCUGAAAACUUCGAGAU [1795-1813](19/19) 419 GCAGAAAAAGAAAACUGGA 922 UCCAGUUUUCUUUUUCUGC [1654-1672](19/19) 420 GGAUAUACGUAGGAGGGGU 923 ACCCCUCCUACGUAUAUCC [1610-1628](19/19) 421 AAGACAAGAACAACUCCAA 924 UUGGAGUUGUUCUUGUCUU [284-302](19/19) 422 AAAGAGCAGUUCAAUGAAG 925 CUUCAUUGAACUGCUCUUU [1572-1590](19/19) 423 UCAACGAAAUGAUGUCCAA 926 UUGGACAUCAUUUCGUUGA [1555-1573](19/19) 424 UAACCUCCCUGUUGUUGAC 927 GUCAACAACAGGGAGGUUA [1535-1553](19/19) 425 CAAAAGCUCUCCAUAUCCC 928 GGGAUAUGGAGAGCUUUUG [1495-1513](19/19) 426 AGAUGAACUUAGGGCAAAA 929 UUUUGCCCUAAGUUCAUCU [1481-1499](19/19) 427 AGAGAUGAACUUAGGGCAA 930 UUGCCCUAAGUUCAUCUCU [1479-1497](19/19) 428 AGCUGGAAAAACAGAAAAA 931 UUUUUCUGUUUUUCCAGCU [253-271](19/19) 429 UCCUAAAACACCAGUACAU 932 AUGUACUGGUGUUUUAGGA [1289-1307](19/19) 430 UGAUGUUUCUGAUCUAUCA 933 UGAUAGAUCAGAAACAUCA [1064-1082](19/19) 431 CCAUUGAUGUUUCUGAUCU 934 AGAUCAGAAACAUCAAUGG [1060-1078](19/19) 432 AGUAUGUUGAAUCAGUAGU 935 ACUACUGAUUCAACAUACU [2096-2114](19/19) 433 UUCUGAGCUAGUUUUUUUG 936 CAAAAAAACUAGCUCAGAA [1952-1970](19/19) 434 UCGAAGUUUUCAGCAUGCU 937 AGCAUGCUGAAAACUUCGA [1798-1816](19/19) 435 AUUCGGGAUAUACGUAGGA 938 UCCUACGUAUAUCCCGAAU [1605-1623](19/19) 436 UGUCCAAAGAGCAGUUCAA 939 UUGAACUGCUCUUUGGACA [1567-1585](19/19) 437 UUCUGGGGAUAUGGUACAA 940 UUGUACCAUAUCCCCAGAA [1310-1328](19/19) 438 CUUCUGGGGAUAUGGUACA 941 UGUACCAUAUCCCCAGAAG [1309-1327](19/19) 439 UGUUUCUGAUCUAUCACUU 942 AAGUGAUAGAUCAGAAACA [1067-1085](19/19) 440 ACCUGCUACUUUAAGCCAU 943 AUGGCUUAAAGUAGCAGGU [1016-1034](19/19) 441 GUUUCAGUCACUUGUUCCU 944 AGGAACAAGUGACUGAAAC [530-548](19/19) 442 CACAUCCAGUCAGAAACCA 945 UGGUUUCUGACUGGAUGUG [384-402](19/19) 443 CUGGUACUAAUAAAGGAUU 946 AAUCCUUUAUUAGUACCAG [2375-2393](19/19) 444 AGUGUCAGUAUGUUGAAUC 947 GAUUCAACAUACUGACACU [2090-2108](19/19) 445 AAACUAGAUUUAGGAGGAU 948 AUCCUCCUAAAUCUAGUUU [1925-1943](19/19) 446 AAAACUAGAUUUAGGAGGA 949 UCCUCCUAAAUCUAGUUUU [1924-1942](19/19) 447 AGCUCAACUUGCAUUAAUU 950 AAUUAAUGCAAGUUGAGCU [1589-1607](19/19) 448 AAGAGCAGUUCAAUGAAGC 951 GCUUCAUUGAACUGCUCUU [1573-1591](19/19) 449 UGAUGUCCAAAGAGCAGUU 952 AACUGCUCUUUGGACAUCA [1564-1582](19/19) 450 GAAAACCCCAUUCACAAAA 953 UUUUGUGAAUGGGGUUUUC [1424-1442](19/19) 451 UGAGCUGGAAAAACAGAAA 954 UUUCUGUUUUUCCAGCUCA [251-269](19/19) 452 UUGAUGUUUCUGAUCUAUC 955 GAUAGAUCAGAAACAUCAA [1063-1081](19/19) 453 UUGCCCACAUUCCCAAAUC 956 GAUUUGGGAAUGUGGGCAA [427-445](19/19) 454 GCACAUCCAGUCAGAAACC 957 GGUUUCUGACUGGAUGUGC [383-401](19/19) 455 GCUGGUACUAAUAAAGGAU 958 AUCCUUUAUUAGUACCAGC [2374-2392](19/19) 456 GGAGUGUCAGUAUGUUGAA 959 UUCAACAUACUGACACUCC [2088-2106](19/19) 457 CCUUUUCUGAGCUAGUUUU 960 AAAACUAGCUCAGAAAAGG [1948-1966](19/19) 458 CUCAAAGAAAAAGGAGAAA 961 UUUCUCCUUUUUCUUUGAG [1731-1749](19/19) 459 AGACAAGAACAACUCCAAA 962 UUUGGAGUUGUUCUUGUCU [285-303](19/19) 460 AUGAUGUCCAAAGAGCAGU 963 ACUGCUCUUUGGACAUCAU [1563-1581](19/19) 461 AUGAGCUGGAAAAACAGAA 964 UUCUGUUUUUCCAGCUCAU [250-268](19/19) 462 GUCCUAAAACACCAGUACA 965 UGUACUGGUGUUUUAGGAC [1288-1306](19/19) 463 AUCAGCAACAGCAUGCCCU 966 AGGGCAUGCUGUUGCUGAU [996-1014](19/19) 464 AAGCCAAACUGACAGAAGU 967 ACUUCUGUCAGUUUGGCUU [763-781](19/19) 465 CGUUUGUAGAUGACAAUGA 968 UCAUUGUCAUCUACAAACG [496-514](19/19) 466 GUUGCCCACAUUCCCAAAU 969 AUUUGGGAAUGUGGGCAAC [426-444](19/19) 467 GCAAACAAGAGAUGGCAAU 970 AUUGCCAUCUCUUGUUUGC [1865-1883](19/19) 468 AUUGCUCAAAGAAAAAGGA 971 UCCUUUUUCUUUGAGCAAU [1727-1745](19/19) 469 AAACCCCAUUCACAAAAGA 972 UCUUUUGUGAAUGGGGUUU [1426-1444](19/19) 470 AAGUGGAAGAGCUAGAUAG 973 CUAUCUAGCUCUUCCACUU [1243-1261](19/19) 471 UCACUAAACACAAGUCCCA 974 UGGGACUUGUGUUUAGUGA [1152-1170](19/19) 472 CGGCAUUUCACUAAACACA 975 UGUGUUUAGUGAAAUGCCG [1145-1163](19/19) 473 UUUAAGCCAUUCACUCUCU 976 AGAGAGUGAAUGGCUUAAA [1025-1043](19/19) 474 CACCUGCUACUUUAAGCCA 977 UGGCUUAAAGUAGCAGGUG [1015-1033](19/19) 475 GGAGGAGCUAUUAUCCAUU 978 AAUGGAUAAUAGCUCCUCC [677-695](19/19) 476 GGGAGGAGCUAUUAUCCAU 979 AUGGAUAAUAGCUCCUCCC [676-694](19/19) 477 UCGCUCAGUUACAACUAGA 980 UCUAGUUGUAACUGAGCGA [325-343](19/19) 478 UGAAGAUGGAAAACCUUAU 981 AUAAGGUUUUCCAUCUUCA [1823-1841](19/19) 479 AAGCUCAACUUGCAUUAAU 982 AUUAAUGCAAGUUGAGCUU [1588-1606](19/19) 480 AAAACCCCAUUCACAAAAG 983 CUUUUGUGAAUGGGGUUUU [1425-1443](19/19) 481 GCUAUGGAGACACACUACU 984 AGUAGUGUGUCUCCAUAGC [1207-1225](19/19) 482 GUAUUUGACUUCAGUCAGC 985 GCUGACUGAAGUCAAAUAC [219-237](19/19) 483 CCUGCUACUUUAAGCCAUU 986 AAUGGCUUAAAGUAGCAGG [1017-1035](19/19) 484 UGAGCCCAGUAUCAGCAAC 987 GUUGCUGAUACUGGGCUCA [986-1004](19/19) 485 GAGAAGUAUUUGACUUCAG 988 CUGAAGUCAAAUACUUCUC [214-232](19/19) 486 UGGAAAAAGAAGUAGGUAA 989 UUACCUACUUCUUUUUCCA [817-835](19/19) 487 GAAGCCAAACUGACAGAAG 990 CUUCUGUCAGUUUGGCUUC [762-780](19/19) 488 CAGAAGCCAAACUGACAGA 991 UCUGUCAGUUUGGCUUCUG [760-778](19/19) 489 GAAAAUGACAAGCUGGUUG 992 CAACCAGCUUGUCAUUUUC [720-738](19/19) 490 AAAAAGCAUUGGAGUGUCA 993 UGACACUCCAAUGCUUUUU [2078-2096](19/19) 491 UAGGAGGAUUUGACCUUUU 994 AAAAGGUCAAAUCCUCCUA [1935-1953](19/19) 492 AGAUGGAAAACCUUAUUCU 995 AGAAUAAGGUUUUCCAUCU [1826-1844](19/19) 493 GAAGCUCAACUUGCAUUAA 996 UUAAUGCAAGUUGAGCUUC [1587-1605](1919) 494 GCUCUCCAUAUCCCAUUCC 997 GGAAUGGGAUAUGGAGAGC [1500-1518](19/19) 495 GGAAAACCCCAUUCACAAA 998 UUUGUGAAUGGGGUUUUCC [1423-1441](19/19) 496 AGAGCACUCACGUGCAUGA 999 UCAUGCACGUGAGUGCUCU [1351-1369](19/19) 497 ACCCUUGUCACCAUCUCAG 1000 CUGAGAUGGUGACAAGGGU [1328-1346](19/19) 498 CUACUUGGCCUCAGUGAUU 1001 AAUCACUGAGGCCAAGUAG [1221-1239](19/19) 499 GUGAACUCAUUAAAUUCAG 1002 CUGAAUUUAAUGAGUUCAC [915-933](19/19) 500 GGAAAAAGAAGUAGGUAAC 1003 GUUACCUACUUCUUUUUCC [818-836](19/19) 501 CCCUGUCGAAAAAAUCAUU 1004 AAUGAUUUUUUCGACAGGG 502 UCCCUGUCGAAAAAAUCAU 1005 AUGAUUUUUUCGACAGGGA 503 CCUGUCGAAAAAAUCAUUA 1006 UAAUGAUUUUUUCGACAGG

TABLE B 21-mers siRNAs specific to Nrf2 gene (SEQ ID NOS: 1007-2006) SEQ ID SEQ ID NO. Sense siRNA NO. Antisense siRNA Human 20149575 1007 UAGGAGGGGUAAGAAUAAAGU  1507 ACUUUAUUCUUACCCCUCCUA [1619-1639](21/21) 1008 AGGGGUAAGAAUAAAGUGGCU 1508 AGCCACUUUAUUCUUACCCCU [1623-1643](21/21) 1009 AAGAAGCCAGAUGUUAAGAAA 1509 UUUCUUAACAUCUGGCUUCUU [1905-1925](21/21) 1010 AAAGGAGAAAAUGACAAAAGC 1510 GCUUUUGUCAUUUUCUCCUUU [1740-1760](21/21) 1011 GAGGGGUAAGAAUAAAGUGGC 1511 GCCACUUUAUUCUUACCCCUC [1622-1642](21/21) 1012 GUAGGAGGGGUAAGAAUAAAG 1512 CUUUAUUCUUACCCCUCCUAC [1618-1638](21/21) 1013 AUAAAGUGGCUGCUCAGAAUU 1513 AAUUCUGAGCAGCCACUUUAU [1633-1653](21/21) 1014 AGGAGGGGUAAGAAUAAAGUG 1514 CACUUUAUUCUUACCCCUCCU [1620-1640](21/21) 1015 GAGAAAGAAUUGCCUGUAAGU 1515 ACUUACAGGCAAUUCUUUCUC [1392-1412](21/21) 1016 UCACUCUCUGAACUUCUAAAU 1516 AUUUAGAAGUUCAGAGAGUGA [1035-1055](21/21) 1017 GACAUUCCCGUUUGUAGAUGA 1517 UCAUCUACAAACGGGAAUGUC [488-508](21/21) 1018 AGAAGCCAGAUGUUAAGAAAA 1518 UUUUCUUAACAUCUGGCUUCU [1906-1926](21/21) 1019 CUCAGAAUUGCAGAAAAAGAA 1519 UUCUUUUUCUGCAAUUCUGAG [1645-1665](21/21) 1020 AAGAAUAAAGUGGCUGCUCAG 1520 CUGAGCAGCCACUUUAUUCUU [1629-1649](21/21) 1021 GGGUAAGAAUAAAGUGGCUGC 1521 GCAGCCACUUUAUUCUUACCC [1625-1645](21/21) 1022 UGAAAAGGAAAGACAAGAACA 1522 UGUUCUUGUCUUUCCUUUUCA [275-295](21/21) 1023 GCCCUCACCUGCUACUUUAAG 1523 CUUAAAGUAGCAGGUGAGGGC [1010-1030](21/21) 1024 GUAGAUGACAAUGAGGUUUCU 1524 AGAAACCUCAUUGUCAUCUAC [501-521](21/21) 1025 GUAAGAAGCCAGAUGUUAAGA 1525 UCUUAACAUCUGGCUUCUUAC [1903-1923](21/21) 1026 AAGUAAGAAGCCAGAUGUUAA 1526 UUAACAUCUGGCUUCUUACUU [1901-1921](21/21) 1027 GGGGUAAGAAUAAAGUGGCUG 1527 CAGCCACUUUAUUCUUACCCC [1624-1644](21/21) 1028 AGAGAAAGAAUUGCCUGUAAG 1528 CUUACAGGCAAUUCUUUCUCU [1391-1411](21/21) 1029 CUCUCUGAACUUCUAAAUGGG 1529 CCCAUUUAGAAGUUCAGAGAG [1038-1058](21/21) 1030 UUACUCAUCUAUACCCUCAAU 1530 AUUGAGGGUAUAGAUGAGUAA [797-817](21/21) 1031 CCAGUCUUCAUUGCUACUAAU 1531 AUUAGUAGCAAUGAAGACUGG [573-593](21/21) 1032 UAGAUGACAAUGAGGUUUCUU 1532 AAGAAACCUCAUUGUCAUCUA [502-522](21/21) 1033 ACAUUCCCGUUUGUAGAUGAC 1533 GUCAUCUACAAACGGGAAUGU [489-509](21/21) 1034 GCAAAACUAACCACUAUGUAC 1534 GUACAUAGUGGUUAGUUUUGC [2270-2290](21/21) 1035 AAAAGUAAGAAGCCAGAUGUU 1535 AACAUCUGGCUUCUUACUUUU [1899-1919](21/21) 1036 CCCAAAAGUAAGAAGCCAGAU 1536 AUCUGGCUUCUUACUUUUGGG [1896-1916](21/21) 1037 UUGCUCAAAGAAAAAGGAGAA 1537 UUCUCCUUUUUCUUUGAGCAA [1728-1748](21/21) 1038 GAACUAGAGCAAGAUUUAGAU 1538 AUCUAAAUCUUGCUCUAGUUC [1683-1703](21/21) 1039 GCUCAGAAUUGCAGAAAAAGA 1539 UCUUUUUCUGCAAUUCUGAGC [1644-1664](21/21) 1040 UUGAAAAGGAAAGACAAGAAC 1540 GUUCUUGUCUUUCCUUUUCAA [274-294](21/21) 1041 AGAACACACCAGAGAAAGAAU 1541 AUUCUUUCUCUGGUGUGUUCU [1381-1401](21/21) 1042 AAAACACCAGUACAUUCUUCU 1542 AGAAGAAUGUACUGGUGUUUU [1293-1313](21/21) 1043 GUCAAACAGAAUGGUCCUAAA 1543 UUUAGGACCAUUCUGUUUGAC [1275-1295](21/21) 1044 AACCAAAACCACCCUGAAAGC 1544 GCUUUCAGGGUGGUUUUGGUU [1098-1118](21/21) 1045 UUCAACCAAAACCACCCUGAA 1545 UUCAGGGUGGUUUUGGUUGAA [1095-1115](21/21) 1046 CACUCUCUGAACUUCUAAAUG 1546 CAUUUAGAAGUUCAGAGAGUG [1036-1056](21/21) 1047 GAGUAAGUCGAGAAGUAUUUG 1547 CAAAUACUUCUCGACUUACUC [205-225](21/21) 1048 GACAAUGAGGUUUCUUCGGCU 1548 AGCCGAAGAAACCUCAUUGUC [507-527](21/21) 1049 UCAGAUGCUUUGUACUUUGAU 1549 AUCAAAGUACAAAGCAUCUGA [444-464](21/21) 1050 GACAUCUGGCUAAAAAGAAAU 1550 AUUUCUUUUUAGCCAGAUGUC [2245-2265](21/21) 1051 GAAGCCAGAUGUUAAGAAAAA 1551 UUUUUCUUAACAUCUGGCUUC [1907-1927](21/21) 1052 CAAAAGUAAGAAGCCAGAUGU 1552 ACAUCUGGCUUCUUACUUUUG [1898-1918](21/21) 1053 AAAACCUUAUUCUCCUAGUGA 1553 UCACUAGGAGAAUAAGGUUUU [1832-1852](21/21) 1054 ACGUAGGAGGGGUAAGAAUAA 1554 UUAUUCUUACCCCUCCUACGU [1616-1636](21/21) 1055 AAGGAAAGACAAGAACAACUC 1555 GAGUUGUUCUUGUCUUUCCUU [279-299](21/21) 1056 CACAAAAGACAAACAUUCAAG 1556 CUUGAAUGUUUGUCUUUUGUG [1436-1456](21/21) 1057 CCCAUUCACAAAAGACAAACA 1557 UGUUUGUCUUUUGUGAAUGGG [1430-1450](21/21) 1058 AACCCCAUUCACAAAAGACAA 1558 UUGUCUUUUGUGAAUGGGGUU [1427-1447](21/21) 1059 ACACCAGAGAAAGAAUUGCCU 1559 AGGCAAUUCUUUCUCUGGUGU [1386-1406](21/21) 1060 AACACACCAGAGAAAGAAUUG 1560 CAAUUCUUUCUCUGGUGUGUU [1383-1403](21/21) 1061 UCAAACAGAAUGGUCCUAAAA 1561 UUUUAGGACCAUUCUGUUUGA [1276-1296](21/21) 1062 UCAGCGACGGAAAGAGUAUGA 1562 UCAUACUCUUUCCGUCGCUGA [233-253](21/21) 1063 GCACAGCAGAAUUCAAUGAUU 1563 AAUCAUUGAAUUCUGCUGUGC [1117-1137](21/21) 1064 GCUUUCAACCAAAACCACCCU 1564 AGGGUGGUUUUGGUUGAAAGC [1092-1112](21/21) 1065 GCCACAGUCAACACAGAUUUU 1565 AAAAUCUGUGUUGACUGUGGC [936-956](21/21) 1066 AACUCAUUAAAUUCAGAUGCC 1566 GGCAUCUGAAUUUAAUGAGUU [918-938](21/21) 1067 GUUGACAGUGAACUCAUUAAA 1567 UUUAAUGAGUUCACUGUCAAC [908-928](21/21) 1068 GCUAUUAUCCAUUCCUGAGUU 1568 AACUCAGGAAUGGAUAAUAGC [683-703](21/21) 1069 AAGUUUGGGAGGAGCUAUUAU 1569 AUAAUAGCUCCUCCCAAACUU [670-690](21/21) 1070 CAAGUUUGGGAGGAGCUAUUA 1570 UAAUAGCUCCUCCCAAACUUG [669-689](21/21) 1071 GUCUUCAUUGCUACUAAUCAG 1571 CUGAUUAGUAGCAAUGAAGAC [576-596](21/21) 1072 GAUGACAAUGAGGUUUCUUCG 1572 CGAAGAAACCUCAUUGUCAUC [504-524](21/21) 1073 AUCAGAUGCUUUGUACUUUGA 1573 UCAAAGUACAAAGCAUCUGAU [443-463](21/21) 1074 GCCCACAUUCCCAAAUCAGAU 1574 AUCUGAUUUGGGAAUGUGGGC [429-449](21/21) 1075 GUGAUGUGAAAUGCUCAUACU 1575 AGUAUGAGCAUUUCACAUCAC [1996-2016](21/21) 1076 AAGCUCCUACUGUGAUGUGAA 1576 UUCACAUCACAGUAGGAGCUU [1985-2005](21/21) 1077 CUACGUGAUGAAGAUGGAAAA 1577 UUUUCCAUCUUCAUCACGUAG [1815-1835](21/21) 1078 AUGCUACGUGAUGAAGAUGGA 1578 UCCAUCUUCAUCACGUAGCAU [1812-1832](21/21) 1079 UGAAAAAACAACUCAGCACCU 1579 AGGUGCUGAGUUGUUUUUUCA [1771-1791](21/21) 1080 UCACCUACUGAAAAAACAACU 1580 AGUUGUUUUUUCAGUAGGUGA [1763-1783](21/21) 1081 GUAGAACUAGAGCAAGAUUUA 1581 UAAAUCUUGCUCUAGUUCUAC [1680-1700](21/21) 1082 UCAACUUGCAUUAAUUCGGGA 1582 UCCCGAAUUAAUGCAAGUUGA [1592-1612](21/21) 1083 AAAGGAAAGACAAGAACAACU 1583 AGUUGUUCUUGUCUUUCCUUU [278-298](21/21) 1084 CCAUUCACAAAAGACAAACAU 1584 AUGUUUGUCUUUUGUGAAUGG [1431-1451](21/21) 1085 CACACCAGAGAAAGAAUUGCC 1585 GGCAAUUCUUUCUCUGGUGUG [1385-1405](21/21) 1086 AACAGAAUGGUCCUAAAACAC 1586 GUGUUUUAGGACCAUUCUGUU [1279-1299](21/21) 1087 GGAAAGAGUAUGAGCUGGAAA 1587 UUUCCAGCUCAUACUCUUUCC [241-261](21/21) 1088 CAGUUGACAGUGAACUCAUUA 1588 UAAUGAGUUCACUGUCAACUG [906-926](21/21) 1089 GGAGUAAGUCGAGAAGUAUUU 1589 AAAUACUUCUCGACUUACUCC [204-224](21/21) 1090 UGGAGUAAGUCGAGAAGUAUU 1590 AAUACUUCUCGACUUACUCCA [203-223](21/21) 1091 UUGGAGUAAGUCGAGAAGUAU 1591 AUACUUCUCGACUUACUCCAA [202-222](21/21) 1092 CUAUUAUCCAUUCCUGAGUUA 1592 UAACUCAGGAAUGGAUAAUAG [684-704](21/21) 1093 GCAACAGGACAUUGAGCAAGU 1593 ACUUGCUCAAUGUCCUGUUGC [653-673](21/21) 1094 GUUGAUUUAGACGGUAUGCAA 1594 UUGCAUACCGUCUAAAUCAAC [636-656](21/21) 1095 CAGUCUUCAUUGCUACUAAUC 1595 GAUUAGUAGCAAUGAAGACUG [574-594](21/21) 1096 UCACAUCGAGAGCCCAGUCUU 1596 AAGACUGGGCUCUCGAUGUGA [560-580](21/21) 1097 AGGACAUGGAUUUGAUUGACA 1597 UGUCAAUCAAAUCCAUGUCCU [157-177](21/21) 1098 UGCCCACAUUCCCAAAUCAGA 1598 UCUGAUUUGGGAAUGUGGGCA [428-448](21/21) 1099 UACUCCCAGGUUGCCCACAUU 1599 AAUGUGGGCAACCUGGGAGUA [417-437](21/21) 1100 AAGAGCUGGUACUAAUAAAGG 1600 CCUUUAUUAGUACCAGCUCUU [2370-2390](21/21) 1101 UAAGAGCUGGUACUAAUAAAG 1601 CUUUAUUAGUACCAGCUCUUA [2369-2389](21/21) 1102 CUGUAUGGACAAAAAAUGGCA 1602 UGCCAUUUUUUGUCCAUACAG [2304-2324](21/21) 1103 GGAUUUGACCUUUUCUGAGCU 1603 AGCUCAGAAAAGGUCAAAUCC [1940-1960](21/21) 1104 AAACCUUAUUCUCCUAGUGAA 1604 UUCACUAGGAGAAUAAGGUUU [1833-1853](21/21) 1105 AAAAAACAACUCAGCACCUUA 1605 UAAGGUGCUGAGUUGUUUUUU [1773-1793](21/21) 1106 CUAGAGCAAGAUUUAGAUCAU 1606 AUGAUCUAAAUCUUGCUCUAG [1686-1706](21/21) 1107 GCAGAAAAAGAAAACUGGAAA 1607 UUUCCAGUUUUCUUUUUCUGC [1654-1674](21/21) 1108 AACUUGCAUUAAUUCGGGAUA 1608 UAUCCCGAAUUAAUGCAAGUU [1594-1614](21/21) 1109 CAACUUGCAUUAAUUCGGGAU 1609 AUCCCGAAUUAAUGCAAGUUG [1593-1613](21/21) 1110 CCAUAUCCCAUUCCCUGUAGA 1610 UCUACAGGGAAUGGGAUAUGG [1505-1525](21/21) 1111 ACAAAAGACAAACAUUCAAGC 1611 GCUUGAAUGUUUGUCUUUUGU [1437-1457](21/21) 1112 ACCCCAUUCACAAAAGACAAA 1612 UUUGUCUUUUGUGAAUGGGGU [1428-1448](21/21) 1113 AAGAAUUGCCUGUAAGUCCUG 1613 CAGGACUUACAGGCAAUUCUU [1396-1416](21/21) 1114 AAAGAAUUGCCUGUAAGUCCU 1614 AGGACUUACAGGCAAUUCUUU [1395-1415](21/21) 1115 GAACACACCAGAGAAAGAAUU 1615 AAUUCUUUCUCUGGUGUGUUC [1382-1402](21/21) 1116 AGAGUAUGAGCUGGAAAAACA 1616 UGUUUUUCCAGCUCAUACUCU [245-265](21/21) 1117 GUAGUCCACAUUUUCUUAAUG 1617 CAUUAAGAAAAUGUGGACUAC [838-858](21/21) 1118 UACUCAUCUAUACCCUCAAUG 1618 CAUUGAGGGUAUAGAUGAGUA [798-818](21/21) 1119 CAGGACAUUGAGCAAGUUUGG 1619 CCAAACUUGCUCAAUGUCCUG [657-677](21/21) 1120 UUCGGCUACGUUUCAGUCACU 1620 AGUGACUGAAACGUAGCCGAA [521-541](21/21) 1121 ACAAUGAGGUUUCUUCGGCUA 1621 UAGCCGAAGAAACCUCAUUGU [508-528](21/21) 1122 GGACAUGGAUUUGAUUGACAU 1622 AUGUCAAUCAAAUCCAUGUCC [158-178](21/21) 1123 UACUGUAUGGACAAAAAAUGG 1623 CCAUUUUUUGUCCAUACAGUA [2302-2322](21/21) 1124 AACAAUUUCUUAGGACACCAU 1624 AUGGUGUCCUAAGAAAUUGUU [2130-2150](21/21) 1125 CUGUAAACAAUUUCUUAGGAC 1625 GUCCUAAGAAAUUGUUUACAG [2125-2145](21/21) 1126 GAAGAUGGAAAACCUUAUUCU 1626 AGAAUAAGGUUUUCCAUCUUC [1824-1844](21/21) 1127 UACGUGAUGAAGAUGGAAAAC 1627 GUUUUCCAUCUUCAUCACGUA [1816-1836](21/21) 1128 AGCACCUUAUAUCUCGAAGUU 1628 AACUUCGAGAUAUAAGGUGCU [1785-1805](21/21) 1129 AAAACAACUCAGCACCUUAUA 1629 UAUAAGGUGCUGAGUUGUUUU [1775-1795](21/21) 1130 GAGCAAGAUUUAGAUCAUUUG 1630 CAAAUGAUCUAAAUCUUGCUC [1689-1709](21/21) 1131 AGAACUAGAGCAAGAUUUAGA 1631 UCUAAAUCUUGCUCUAGUUCU [1682-1702](21/21) 1132 UUGCAGAAAAAGAAAACUGGA 1632 UCCAGUUUUCUUUUUCUGCAA [1652-1672](21/21) 1133 UCCAUAUCCCAUUCCCUGUAG 1633 CUACAGGGAAUGGGAUAUGGA [1504-1524](21/21) 1134 CCCCAUUCACAAAAGACAAAC 1634 GUUUGUCUUUUGUGAAUGGGG [1429-1449](21/21) 1135 GAGAACACACCAGAGAAAGAA 1635 UUCUUUCUCUGGUGUGUUCUC [1380-1400](21/21) 1136 CAGUACAUUCUUCUGGGGAUA 1636 UAUCCCCAGAAGAAUGUACUG [1300-1320](21/21) 1137 ACGGAAAGAGUAUGAGCUGGA 1637 UCCAGCUCAUACUCUUUCCGU [239-259](21/21) 1138 CUGACUCCGGCAUUUCACUAA 1638 UUAGUGAAAUGCCGGAGUCAG [1138-1158](21/21) 1139 AGCACAGCAGAAUUCAAUGAU 1639 AUCAUUGAAUUCUGCUGUGCU [1116-1136](21/21) 1140 UAAAUUCAGAUGCCACAGUCA 1640 UGACUGUGGCAUCUGAAUUUA [925-945](21/21) 1141 CUCAUUAAAUUCAGAUGCCAC 1641 GUGGCAUCUGAAUUUAAUGAG [920-940](21/21) 1142 ACUGUAGUCCACAUUUUCUUA 1642 UAAGAAAAUGUGGACUACAGU [835-855](21/21) 1143 UCAAUGGAAAAAGAAGUAGGU 1643 ACCUACUUCUUUUUCCAUUGA [813-833](21/21) 1144 GAGCUAUUAUCCAUUCCUGAG 1644 CUCAGGAAUGGAUAAUAGCUC [681-701](21/21) 1145 AACAGGACAUUGAGCAAGUUU 1645 AAACUUGCUCAAUGUCCUGUU [655-675](21/21) 1146 UUAGACGGUAUGCAACAGGAC 1646 GUCCUGUUGCAUACCGUCUAA [642-662](21/21) 1147 UUUAGACGGUAUGCAACAGGA 1647 UCCUGUUGCAUACCGUCUAAA [641-661](21/21) 1148 UGAGGUUUCUUCGGCUACGUU 1648 AACGUAGCCGAAGAAACCUCA [512-532](21/21) 1149 AUGAGGUUUCUUCGGCUACGU 1649 ACGUAGCCGAAGAAACCUCAU [511-531](21/21) 1150 UGCUUUGUACUUUGAUGACUG 1650 CAGUCAUCAAAGUACAAAGCA [449-469](21/21) 1151 AGAUGCUUUGUACUUUGAUGA 1651 UCAUCAAAGUACAAAGCAUCU [446-466](21/21) 1152 CAGCAGGACAUGGAUUUGAUU 1652 AAUCAAAUCCAUGUCCUGCUG [153-173](21/21) 1153 UUACAACUAGAUGAAGAGACA 1653 UGUCUCUUCAUCUAGUUGUAA [333-353](21/21) 1154 AGAGCUGGUACUAAUAAAGGA 1654 UCCUUUAUUAGUACCAGCUCU [2371-2391](21/21) 1155 UGACAUCUGGCUAAAAAGAAA 1655 UUUCUUUUUAGCCAGAUGUCA [2244-2264](21/21) 1156 CUAGUUUCUGUGUAAGUGUAA 1656 UUACACUUACACAGAAACUAG [2156-2176](21/21) 1157 GGCUAGUUUCUGUGUAAGUGU 1657 ACACUUACACAGAAACUAGCC [2154-2174](21/21) 1158 UGGGCUAGUUUCUGUGUAAGU 1658 ACUUACACAGAAACUAGCCCA [2152-2172](21/21) 1159 AAACAAUUUCUUAGGACACCA 1659 UGGUGUCCUAAGAAAUUGUUU [2129-2149](21/21) 1160 GAGUGUCAGUAUGUUGAAUCA 1660 UGAUUCAACAUACUGACACUC [2089-2109](21/21) 1161 UACUAAAAGCUCCUACUGUGA 1661 UCACAGUAGGAGCUUUUAGUA [1979-1999](21/21) 1162 UGCUACGUGAUGAAGAUGGAA 1662 UUCCAUCUUCAUCACGUAGCA [1813-1833](21/21) 1163 GCACCUUAUAUCUCGAAGUUU 1663 AAACUUCGAGAUAUAAGGUGC [1786-1806](21/21) 1164 CAGCACCUUAUAUCUCGAAGU 1664 ACUUCGAGAUAUAAGGUGCUG [1784-1804](21/21) 1165 CACCUACUGAAAAAACAACUC 1665 GAGUUGUUUUUUCAGUAGGUG [1764-1784](21/21) 1166 AACUAGAGCAAGAUUUAGAUC 1666 GAUCUAAAUCUUGCUCUAGUU [1684-1704](21/21) 1167 UAGUAGAACUAGAGCAAGAUU 1667 AAUCUUGCUCUAGUUCUACUA [1678-1698](21/21) 1168 GGGAUAUACGUAGGAGGGGUA 1668 UACCCCUCCUACGUAUAUCCC [1609-1629](21/21) 1169 UGAGAACACACCAGAGAAAGA 1669 UCUUUCUCUGGUGUGUUCUCA [1379-1399](21/21) 1170 AAAGAGUAUGAGCUGGAAAAA 1670 UUUUUCCAGCUCAUACUCUUU [243-263](21/21) 1171 CGGAAAGAGUAUGAGCUGGAA 1671 UUCCAGCUCAUACUCUUUCCG [240-260](21/21) 1172 UGCCCCUGGAAGUGUCAAACA 1672 UGUUUGACACUUCCAGGGGCA [1262-1282](21/21) 1173 AUAGUGCCCCUGGAAGUGUCA 1673 UGACACUUCCAGGGGCACUAU [1258-1278](21/21) 1174 AGCGACGGAAAGAGUAUGAGC 1674 GCUCAUACUCUUUCCGUCGCU [235-255](21/21) 1175 CAGCGACGGAAAGAGUAUGAG 1675 CUCAUACUCUUUCCGUCGCUG [234-254](21/21) 1176 GGGCCCAUUGAUGUUUCUGAU 1676 AUCAGAAACAUCAAUGGGCCC [1056-1076](21/21) 1177 CUGUAGUCCACAUUUUCUUAA 1677 UUAAGAAAAUGUGGACUACAG [836-856](21/21) 1178 CUCAAUGGAAAAAGAAGUAGG 1678 CCUACUUCUUUUUCCAUUGAG [812-832](21/21) 1179 GAUCUUGGAGUAAGUCGAGAA 1679 UUCUCGACUUACUCCAAGAUC [198-218](21/21) 1180 CUUUGGAGGCAAGAUAUAGAU 1680 AUCUAUAUCUUGCCUCCAAAG [180-200](21/21) 1181 GACAUUGAGCAAGUUUGGGAG 1681 CUCCCAAACUUGCUCAAUGUC [660-680](21/21) 1182 GGACAUUGAGCAAGUUUGGGA 1682 UCCCAAACUUGCUCAAUGUCC [659-679](21/21) 1183 GUCACCUGAAACUUCUGUUGC 1683 GCAACAGAAGUUUCAGGUGAC [602-622](21/21) 1184 GCCCAGUCUUCAUUGCUACUA 1684 UAGUAGCAAUGAAGACUGGGC [571-591](21/21) 1185 UCGAGAGCCCAGUCUUCAUUG 1685 CAAUGAAGACUGGGCUCUCGA [565-585](21/21) 1186 UCGGCUACGUUUCAGUCACUU 1686 AAGUGACUGAAACGUAGCCGA [522-542](21/21) 1187 GAGGUUUCUUCGGCUACGUUU 1687 AAACGUAGCCGAAGAAACCUC [513-533](21/21) 1188 UUGUACUUUGAUGACUGCAUG 1688 CAUGCAGUCAUCAAAGUACAA [453-473](21/21) 1189 AUGCUUUGUACUUUGAUGACU 1689 AGUCAUCAAAGUACAAAGCAU [448-468](21/21) 1190 GAUGCUUUGUACUUUGAUGAC 1690 GUCAUCAAAGUACAAAGCAUC [447-467](21/21) 1191 CUAGAUGAAGAGACAGGUGAA 1691 UUCACCUGUCUCUUCAUCUAG [339-359](21/21) 1192 GUAUGGACAAAAAAUGGCAUU 1692 AAUGCCAUUUUUUGUCCAUAC [2306-2326](21/21) 1193 UGUAUGGACAAAAAAUGGCAU 1693 AUGCCAUUUUUUGUCCAUACA [2305-2325](21/21) 1194 ACAAUUUCUUAGGACACCAUU 1694 AAUGGUGUCCUAAGAAAUUGU [2131-2151](21/21) 1195 UGAUGUGAAAUGCUCAUACUU 1695 AAGUAUGAGCAUUUCACAUCA [1997-2017](21/21) 1196 UCCUACUGUGAUGUGAAAUGC 1696 GCAUUUCACAUCACAGUAGGA [1989-2009](21/21) 1197 UCCUAGUGAAUACUCCCUGCA 1697 UGCAGGGAGUAUUCACUAGGA [1844-1864](21/21) 1198 ACACGGUCCACAGCUCAUCAU 1698 AUGAUGAGCUGUGGACCGUGU [95-115](21/21) 1199 GAAGUGUCAAACAGAAUGGUC 1699 GACCAUUCUGUUUGACACUUC [1270-1290](21/21) 1200 UGGAAGUGUCAAACAGAAUGG 1700 CCAUUCUGUUUGACACUUCCA [1268-1288](21/21) 1201 GUGAUUCUGAAGUGGAAGAGC 1701 GCUCUUCCACUUCAGAAUCAC [1234-1254](21/21) 1202 GAAAGCACAGCAGAAUUCAAU 1702 AUUGAAUUCUGCUGUGCUUUC [1113-1133](21/21) 1203 CACAGUCAACACAGAUUUUGG 1703 CCAAAAUCUGUGUUGACUGUG [938-958](21/21) 1204 CAGAAGCCAAACUGACAGAAG 1704 CUUCUGUCAGUUUGGCUUCUG [760-780](21/21) 1205 ACCUGAAACUUCUGUUGCUCA 1705 UGAGCAACAGAAGUUUCAGGU [605-625](21/21) 1206 UUGACAUACUUUGGAGGCAAG 1706 CUUGCCUCCAAAGUAUGUCAA [172-192](21/21) 1207 UUGAUUGACAUACUUUGGAGG 1707 CCUCCAAAGUAUGUCAAUCAA [168-188](21/21) 1208 GUCACUUGUUCCUGAUAUUCC 1708 GGAAUAUCAGGAACAAGUGAC [536-556](21/21) 1209 GCUUUGUACUUUGAUGACUGC 1709 GCAGUCAUCAAAGUACAAAGC [450-470](21/21) 1210 CAGGACAUGGAUUUGAUUGAC 1710 GUCAAUCAAAUCCAUGUCCUG [156-176](21/21) 1211 CAGUUACAACUAGAUGAAGAG 1711 CUCUUCAUCUAGUUGUAACUG [330-350](21/21) 1212 GAUAUGACAUCUGGCUAAAAA 1712 UUUUUAGCCAGAUGUCAUAUC [2240-2260](21/21) 1213 CAGUAUGUUGAAUCAGUAGUU 1713 AACUACUGAUUCAACAUACUG [2095-2115](21/21) 1214 AGCAUUGGAGUGUCAGUAUGU 1714 ACAUACUGACACUCCAAUGCU [2082-2102](21/21) 1215 AAAGCAUUGGAGUGUCAGUAU 1715 AUACUGACACUCCAAUGCUUU [2080-2100](21/21) 1216 CCUACUGUGAUGUGAAAUGCU 1716 AGCAUUUCACAUCACAGUAGG [1990-2010](21/21) 1217 UGACCUUUUCUGAGCUAGUUU 1717 AAACUAGCUCAGAAAAGGUCA [1945-1965](21/21) 1218 GAUUUGACCUUUUCUGAGCUA 1718 UAGCUCAGAAAAGGUCAAAUC [1941-1961](21/21) 1219 CAGCAUGCUACGUGAUGAAGA 1719 UCUUCAUCACGUAGCAUGCUG [1808-1828](21/21) 1220 CACCUUAUAUCUCGAAGUUUU 1720 AAAACUUCGAGAUAUAAGGUG [1787-1807](21/21) 1221 UCAGCACCUUAUAUCUCGAAG 1721 CUUCGAGAUAUAAGGUGCUGA [1783-1803](21/21) 1222 ACUCAGCACCUUAUAUCUCGA 1722 UCGAGAUAUAAGGUGCUGAGU [1781-1801](21/21) 1223 AUAGUAGAACUAGAGCAAGAU 1723 AUCUUGCUCUAGUUCUACUAU [1677-1697](21/21) 1224 GGAUAUACGUAGGAGGGGUAA 1724 UUACCCCUCCUACGUAUAUCC [1610-1630](21/21) 1225 AAAGCUCUCCAUAUCCCAUUC 1725 GAAUGGGAUAUGGAGAGCUUU [1497-1517](21/21) 1226 GCAAAAGCUCUCCAUAUCCCA 1726 UGGGAUAUGGAGAGCUUUUGC [1494-1514](21/21) 1227 AGAGAUGAACUUAGGGCAAAA 1727 UUUUGCCCUAAGUUCAUCUCU [1479-1499](21/21) 1228 CAAGAGAUGAACUUAGGGCAA 1728 UUGCCCUAAGUUCAUCUCUUG [1477-1497](21/21) 1229 UCACAAGAGAUGAACUUAGGG 1729 CCCUAAGUUCAUCUCUUGUGA [1474-1494](21/21) 1230 GGAAGUGUCAAACAGAAUGGU 1730 ACCAUUCUGUUUGACACUUCC [1269-1289](21/21) 1231 CCUGGAAGUGUCAAACAGAAU 1731 AUUCUGUUUGACACUUCCAGG [1266-1286](21/21) 1232 GAUUCUGAAGUGGAAGAGCUA 1732 UAGCUCUUCCACUUCAGAAUC [1236-1256](21/21) 1233 GCGACGGAAAGAGUAUGAGCU 1733 AGCUCAUACUCUUUCCGUCGC [236-256](21/21) 1234 UUCUGAUCUAUCACUUUGCAA 1734 UUGCAAAGUGAUAGAUCAGAA [1070-1090](21/21) 1235 AUCUUGGAGUAAGUCGAGAAG 1735 CUUCUCGACUUACUCCAAGAU [199-219](21/21) 1236 CAACAGGACAUUGAGCAAGUU 1736 AACUUGCUCAAUGUCCUGUUG [654-674](21/21) 1237 UCACCUGAAACUUCUGUUGCU 1737 AGCAACAGAAGUUUCAGGUGA [603-623](21/21) 1238 AUCGAGAGCCCAGUCUUCAUU 1738 AAUGAAGACUGGGCUCUCGAU [564-584](21/21) 1239 UACGUUUCAGUCACUUGUUCC 1739 GGAACAAGUGACUGAAACGUA [527-547](21/21) 1240 GCUACGUUUCAGUCACUUGUU 1740 AACAAGUGACUGAAACGUAGC [525-545](21/21) 1241 GGCUACGUUUCAGUCACUUGU 1741 ACAAGUGACUGAAACGUAGCC [524-544](21/21) 1242 CGGCUACGUUUCAGUCACUUG 1742 CAAGUGACUGAAACGUAGCCG [523-543](21/21) 1243 UUUCUUCGGCUACGUUUCAGU 1743 ACUGAAACGUAGCCGAAGAAA [517-537](21/21) 1244 GACAUGGAUUUGAUUGACAUA 1744 UAUGUCAAUCAAAUCCAUGUC [159-179](21/21) 1245 AUAUGACAUCUGGCUAAAAAG 1745 CUUUUUAGCCAGAUGUCAUAU [2241-2261](21/21) 1246 UUGGGCUAGUUUCUGUGUAAG 1746 CUUACACAGAAACUAGCCCAA [2151-2171](21/21) 1247 GUCAGUAUGUUGAAUCAGUAG 1747 CUACUGAUUCAACAUACUGAC [2093-2113](21/21) 1248 UGUCAGUAUGUUGAAUCAGUA 1748 UACUGAUUCAACAUACUGACA [2092-2112](21/21) 1249 GUGUCAGUAUGUUGAAUCAGU 1749 ACUGAUUCAACAUACUGACAC [2091-2111](21/21) 1250 UUGGAGUGUCAGUAUGUUGAA 1750 UUCAACAUACUGACACUCCAA [2086-2106](21/21) 1251 UACUGUGAUGUGAAAUGCUCA 1751 UGAGCAUUUCACAUCACAGUA [1992-2012](21/21) 1252 ACCUUUUCUGAGCUAGUUUUU 1752 AAAAACUAGCUCAGAAAAGGU [1947-1967](21/21) 1253 UUUGACCUUUUCUGAGCUAGU 1753 ACUAGCUCAGAAAAGGUCAAA [1943-1963](21/21) 1254 UGCAGCAAACAAGAGAUGGCA 1754 UGCCAUCUCUUGUUUGCUGCA [1861-1881](21/21) 1255 CUCCUAGUGAAUACUCCCUGC 1755 GCAGGGAGUAUUCACUAGGAG [1843-1863](21/21) 1256 GCAUGCUACGUGAUGAAGAUG 1756 CAUCUUCAUCACGUAGCAUGC [1810-1830](21/21) 1257 ACCUUAUAUCUCGAAGUUUUC 1757 GAAAACUUCGAGAUAUAAGGU [1788-1808](21/21) 1258 UUAAUUCGGGAUAUACGUAGG 1758 CCUACGUAUAUCCCGAAUUAA [1602-1622](21/21) 1259 CAACGAAAUGAUGUCCAAAGA 1759 UCUUUGGACAUCAUUUCGUUG [1556-1576](21/21) 1260 AACCUCCCUGUUGUUGACUUC 1760 GAAGUCAACAACAGGGAGGUU [1536-1556](21/21) 1261 GAGAUGAACUUAGGGCAAAAG 1761 CUUUUGCCCUAAGUUCAUCUC [1480-1500](21/21) 1262 ACAAGAGAUGAACUUAGGGCA 1762 UGCCCUAAGUUCAUCUCUUGU [1476-1496](21/21) 1263 AAAGCACAGCAGAAUUCAAUG 1763 CAUUGAAUUCUGCUGUGCUUU [1114-1134](21/21) 1264 GAUGUUUCUGAUCUAUCACUU 1764 AAGUGAUAGAUCAGAAACAUC [1065-1085](21/21) 1265 ACAGUCAACACAGAUUUUGGU 1765 ACCAAAAUCUGUGUUGACUGU [939-959](21/21) 1266 UGAGGAUUCCUUCAGCAGCAU 1766 AUGCUGCUGAAGGAAUCCUCA [863-883](21/21) 1267 UGACAUACUUUGGAGGCAAGA 1767 UCUUGCCUCCAAAGUAUGUCA [173-193](21/21) 1268 CAUGGAUUUGAUUGACAUACU 1768 AGUAUGUCAAUCAAAUCCAUG [161-181](21/21) 1269 GCAGGACAUGGAUUUGAUUGA 1769 UCAAUCAAAUCCAUGUCCUGC [155-175](21/21) 1270 AUGAAGAGACAGGUGAAUUUC 1770 GAAAUUCACCUGUCUCUUCAU [343-363](21/21) 1271 AGAUGAAGAGACAGGUGAAUU 1771 AAUUCACCUGUCUCUUCAUCU [341-361](21/21) 1272 UUGCAAAACUAACCACUAUGU 1772 ACAUAGUGGUUAGUUUUGCAA [2268-2288](21/21) 1273 AUGACAUCUGGCUAAAAAGAA 1773 UUCUUUUUAGCCAGAUGUCAU [2243-2263](21/21) 1274 GGGCUAGUUUCUGUGUAAGUG 1774 CACUUACACAGAAACUAGCCC [2153-2173](21/21) 1275 UUUGGGCUAGUUUCUGUGUAA 1775 UUACACAGAAACUAGCCCAAA [2150-2170](21/21) 1276 GGAGUGUCAGUAUGUUGAAUC 1776 GAUUCAACAUACUGACACUCC [2088-2108](21/21) 1277 UGUGAUGUGAAAUGCUCAUAC 1777 GUAUGAGCAUUUCACAUCACA [1995-2015](21/21) 1278 CUCAGCACCUUAUAUCUCGAA 1778 UUCGAGAUAUAAGGUGCUGAG [1782-1802](21/21) 1279 UAAUUCGGGAUAUACGUAGGA 1779 UCCUACGUAUAUCCCGAAUUA [1603-1623](21/21) 1280 AAAGACAAGAACAACUCCAAA 1780 UUUGGAGUUGUUCUUGUCUUU [283-303](21/21) 1281 GAAAGACAAGAACAACUCCAA 1781 UUGGAGUUGUUCUUGUCUUUC [282-302](21/21) 1282 CGAAAUGAUGUCCAAAGAGCA 1782 UGCUCUUUGGACAUCAUUUCG [1559-1579](21/21) 1283 AACGAAAUGAUGUCCAAAGAG 1783 CUCUUUGGACAUCAUUUCGUU [1557-1577](21/21) 1284 ACCUCCCUGUUGUUGACUUCA 1784 UGAAGUCAACAACAGGGAGGU [1537-1557](21/21) 1285 AUCAUUAACCUCCCUGUUGUU 1785 AACAACAGGGAGGUUAAUGAU [1530-1550](21/21) 1286 UUUCUGAUCUAUCACUUUGCA 1786 UGCAAAGUGAUAGAUCAGAAA [1069-1089](21/21) 1287 CACUUGUUCCUGAUAUUCCCG 1787 CGGGAAUAUCAGGAACAAGUG [538-558](21/21) 1288 GAUUUGAUUGACAUACUUUGG 1788 CCAAAGUAUGUCAAUCAAAUC [165-185](21/21) 1289 ACGUUUCAGUCACUUGUUCCU 1789 AGGAACAAGUGACUGAAACGU [528-548](21/21) 1290 ACAGGUGAAUUUCUCCCAAUU 1790 AAUUGGGAGAAAUUCACCUGU [351-371](21/21) 1291 GAGCUGGUACUAAUAAAGGAU 1791 AUCCUUUAUUAGUACCAGCUC [2372-2392](21/21) 1292 CUGUGAUGUGAAAUGCUCAUA 1792 UAUGAGCAUUUCACAUCACAG [1994-2014](21/21) 1293 GAAAAACUAGAUUUAGGAGGA 1793 UCCUCCUAAAUCUAGUUUUUC [1922-1942](21/21) 1294 GCUCAAAGAAAAAGGAGAAAA 1794 UUUUCUCCUUUUUCUUUGAGC [1730-1750](21/21) 1295 AGACAAGAACAACUCCAAAAG 1795 CUUUUGGAGUUGUUCUUGUCU [285-305](21/21) 1296 UUCGGGAUAUACGUAGGAGGG 1796 CCCUCCUACGUAUAUCCCGAA [1606-1626](21/21) 1297 AUGUCCAAAGAGCAGUUCAAU 1797 AUUGAACUGCUCUUUGGACAU [1566-1586](21/21) 1298 UGAUGUCCAAAGAGCAGUUCA 1798 UGAACUGCUCUUUGGACAUCA [1564-1584](21/21) 1299 GAGCUGGAAAAACAGAAAAAA 1799 UUUUUUCUGUUUUUCCAGCUC [252-272](21/21) 1300 CCGGCAUUUCACUAAACACAA 1800 UUGUGUUUAGUGAAAUGCCGG [1144-1164](21/21) 1301 UUGAUGUUUCUGAUCUAUCAC 1801 GUGAUAGAUCAGAAACAUCAA [1063-1083](21/21) 1302 CCAUUGAUGUUUCUGAUCUAU 1802 AUAGAUCAGAAACAUCAAUGG [1060-1080](21/21) 1303 CCCAUUGAUGUUUCUGAUCUA 1803 UAGAUCAGAAACAUCAAUGGG [1059-1079](21/21) 1304 GGCCCAUUGAUGUUUCUGAUC 1804 GAUCAGAAACAUCAAUGGGCC [1057-1077](21/21) 1305 AAGAAGUAGGUAACUGUAGUC 1805 GACUACAGUUACCUACUUCUU [823-843](21/21) 1306 GACCUUUUCUGAGCUAGUUUU 1806 AAAACUAGCUCAGAAAAGGUC [1946-1966](21/21) 1307 AUCUCGAAGUUUUCAGCAUGC 1807 GCAUGCUGAAAACUUCGAGAU [1795-1815](21/21) 1308 AAGACAAGAACAACUCCAAAA 1808 UUUUGGAGUUGUUCUUGUCUU [284-304](21/21) 1309 GAAAACCCCAUUCACAAAAGA 1809 UCUUUUGUGAAUGGGGUUUUC [1424-1444](21/21) 1310 GAUAUGGUACAACCCUUGUCA 1810 UGACAAGGGUUGUACCAUAUC [1317-1337](21/21) 1311 GAAGUGGAAGAGCUAGAUAGU 1811 ACUAUCUAGCUCUUCCACUUC [1242-1262](21/21) 1312 GUUUCUGAUCUAUCACUUUGC 1812 GCAAAGUGAUAGAUCAGAAAC [1068-1088](21/21) 1313 GCCCAUUGAUGUUUCUGAUCU 1813 AGAUCAGAAACAUCAAUGGGC [1058-1078](21/21) 1314 UUGCCCACAUUCCCAAAUCAG 1814 CUGAUUUGGGAAUGUGGGCAA [427-447](21/21) 1315 GUUGCCCACAUUCCCAAAUCA 1815 UGAUUUGGGAAUGUGGGCAAC [426-446](21/21) 1316 GCAGCAAACAAGAGAUGGCAA 1816 UUGCCAUCUCUUGUUUGCUGC [1862-1882](21/21) 1317 UGCUCAAAGAAAAAGGAGAAA 1817 UUUCUCCUUUUUCUUUGAGCA [1729-1749](21/21) 1318 GAAGCUCAACUUGCAUUAAUU 1818 AAUUAAUGCAAGUUGAGCUUC [1587-1607](21/21) 1319 CAAAGAGCAGUUCAAUGAAGC 1819 GCUUCAUUGAACUGCUCUUUG [1571-1591](21/21) 1320 GGAAAACCCCAUUCACAAAAG 1820 CUUUUGUGAAUGGGGUUUUCC [1423-1443](21/21) 1321 UGAGCUGGAAAAACAGAAAAA 1821 UUUUUCUGUUUUUCCAGCUCA [251-271](21/21) 1322 GCCCAGUAUCAGCAACAGCAU 1822 AUGCUGUUGCUGAUACUGGGC [989-1009](21/21) 1323 UAGGUAACUGUAGUCCACAUU 1823 AAUGUGGACUACAGUUACCUA [829-849](21/21) 1324 CAUUCCUGAGUUACAGUGUCU 1824 AGACACUGUAACUCAGGAAUG [692-712](21/21) 1325 CCGUUUGUAGAUGACAAUGAG 1825 CUCAUUGUCAUCUACAAACGG [495-515](21/21) 1326 GCUGGUACUAAUAAAGGAUUA 1826 UAAUCCUUUAUUAGUACCAGC [2374-2394](21/21) 1327 AGGACACCAUUUGGGCUAGUU 1827 AACUAGCCCAAAUGGUGUCCU [2141-2161](21/21) 1328 GCUCUCCAUAUCCCAUUCCCU 1828 AGGGAAUGGGAUAUGGAGAGC [1500-1520](21/21) 1329 UGGGCCCAUUGAUGUUUCUGA 1829 UCAGAAACAUCAAUGGGCCCA [1055-1075](21/21) 1330 ACUUUAAGCCAUUCACUCUCU 1830 AGAGAGUGAAUGGCUUAAAGU [1023-1043](21/21) 1331 CACCUGCUACUUUAAGCCAUU 1831 AAUGGCUUAAAGUAGCAGGUG [1015-1035](21/21) 1332 CUCACCUGCUACUUUAAGCCA 1832 UGGCUUAAAGUAGCAGGUGAG [1013-1033](21/21) 1333 UGAAAAUGACAAGCUGGUUGA 1833 UCAACCAGCUUGUCAUUUUCA [719-739](21/21) 1334 AGCUGGUACUAAUAAAGGAUU 1834 AAUCCUUUAUUAGUACCAGCU [2373-2393](21/21) 1335 AUGCAAAAUCAUAGCCAAAAC 1835 GUUUUGGCUAUGAUUUUGCAU [2031-2051](21/21) 1336 CAAACAAGAGAUGGCAAUGUU 1836 AACAUUGCCAUCUCUUGUUUG [1866-1886](21/21) 1337 GAAAAUGACAAAAGCCUUCAC 1837 GUGAAGGCUUUUGUCAUUUUC [1746-1766](21/21) 1338 AAAACCCCAUUCACAAAAGAC 1838 GUCUUUUGUGAAUGGGGUUUU [1425-1445](21/21) 1339 AUGAGCUGGAAAAACAGAAAA 1839 UUUUCUGUUUUUCCAGCUCAU [250-270](21/21) 1340 CUGAAGUGGAAGAGCUAGAUA 1840 UAUCUAGCUCUUCCACUUCAG [1240-1260](21/21) 1341 ACUCCGGCAUUUCACUAAACA 1841 UGUUUAGUGAAAUGCCGGAGU [1141-1161](21/21) 1342 CCAUUCACUCUCUGAACUUCU 1842 AGAAGUUCAGAGAGUGAAUGG [1031-1051](21/21) 1343 AGCCAUUCACUCUCUGAACUU 1843 AAGUUCAGAGAGUGAAUGGCU [1029-1049](21/21) 1344 CUGCUACUUUAAGCCAUUCAC 1844 GUGAAUGGCUUAAAGUAGCAG [1018-1038](21/21) 1345 UCACCUGCUACUUUAAGCCAU 1845 AUGGCUUAAAGUAGCAGGUGA [1014-1034](21/21) 1346 UGCCCUCACCUGCUACUUUAA 1846 UUAAAGUAGCAGGUGAGGGCA [1009-1029](21/21) 1347 AUGCCCUCACCUGCUACUUUA 1847 UAAAGUAGCAGGUGAGGGCAU [1008-1028](21/21) 1348 AUGGAAAAAGAAGUAGGUAAC 1848 GUUACCUACUUCUUUUUCCAU [816-836](21/21) 1349 GAAGCCAAACUGACAGAAGUU 1849 AACUUCUGUCAGUUUGGCUUC [762-782](21/21) 1350 UUAAAAAGCAUUGGAGUGUCA 1850 UGACACUCCAAUGCUUUUUAA [2076-2096](21/21) 1351 GGCAAUGUUUUCCUUGUUCCC 1851 GGGAACAAGGAAAACAUUGCC [1878-1898](21/21) 1352 ACAACUCAGCACCUUAUAUCU 1852 AGAUAUAAGGUGCUGAGUUGU [1778-1798](21/21) 1353 GGCUGCUCAGAAUUGCAGAAA 1853 UUUCUGCAAUUCUGAGCAGCC [1640-1660](21/21) 1354 AGCUCUCCAUAUCCCAUUCCC 1854 GGGAAUGGGAUAUGGAGAGCU [1499-1519](21/21) 1355 AAGCUCUCCAUAUCCCAUUCC 1855 GGAAUGGGAUAUGGAGAGCUU [1498-1518](21/21) 1356 AAGACAAACAUUCAAGCCGCU 1856 AGCGGCUUGAAUGUUUGUCUU [1441-1461](21/21) 1357 AAACCCCAUUCACAAAAGACA 1857 UGUCUUUUGUGAAUGGGGUUU [1426-1446](21/21) 1358 CGGCAUUUCACUAAACACAAG 1858 CUUGUGUUUAGUGAAAUGCCG [1145-1165](21/21) 1359 CCCGUUUGUAGAUGACAAUGA 1859 UCAUUGUCAUCUACAAACGGG [494-514](21/21) 1360 UCCCGUUUGUAGAUGACAAUG 1860 CAUUGUCAUCUACAAACGGGA [493-513](21/21) 1361 AUCUGCCAACUACUCCCAGGU 1861 ACCUGGGAGUAGUUGGCAGAU [407-427](21/21) 1362 GCCAGAUGUUAAGAAAAACUA 1862 UAGUUUUUCUUAACAUCUGGC [1910-1930](21/21) 1363 AAAUGACAAAAGCCUUCACCU 1863 AGGUGAAGGCUUUUGUCAUUU [1748-1768](21/21) 1364 CUGCUCAGAAUUCCAGAAAAA 1864 UUUUUCUGCAAUUCUGAGCAG [1642-1662](21/21) 1365 AAGCUCAACUUGCAUUAAUUC 1865 GAAUUAAUGCAAGUUGAGCUU [1588-1608](21/21) 1366 GUCAUCGGAAAACCCCAUUCA 1866 UGAAUGGGGUUUUCCGAUGAC [1417-1437](21/21) 1367 CCCCAACCAGUUGACAGUGAA 1867 UUCACUGUCAACUGGUUGGGG [899-919](21/21) 1368 GAAAAAGAAGUAGGUAACUGU 1868 ACAGUUACCUACUUCUUUUUC [819-839](21/21) 1369 UGGAAAAAGAAGUAGGUAACU 1869 AGUUACCUACUUCUUUUUCCA [817-837](21/21) 1370 CAAGUCCAGAAGCCAAACUGA 1870 UCAGUUUGGCUUCUGGACUUG [754-774](21/21) 1371 UGGGAGGAGCUAUUAUCCAUU 1871 AAUGGAUAAUAGCUCCUCCCA [675-695](21/21) 1372 UGCAAAACUAACCACUAUGUA 1872 UACAUAGUGGUUAGUUUUGCA [2269-2289](21/21) 1373 AUGGAAAACCUUAUUCUCCUA 1873 UAGGAGAAUAAGGUUUUCCAU [1828-1848](21/21) 1374 AGCCUUCACCUACUGAAAAAA 1874 UUUUUUCAGUAGGUGAAGGCU [1758-1778](21/21) 1375 AAGGAGAAAAUGACAAAAGCC 1875 GGCUUUUGUCAUUUUCUCCUU [1741-1761](21/21) 1376 AAAGACAAACAUUCAAGCCGC 1876 GCGGCUUGAAUGUUUGUCUUU [1440-1460](21/21) 1377 UAAGUCCUGGUCAUCGGAAAA 1877 UUUUCCGAUGACCAGGACUUA [1408-1428](21/21) 1378 UAUGAGCUGGAAAAACAGAAA 1878 UUUCUGUUUUUCCAGCUCAUA [249-269](21/21) 1379 ACAACCCUUGUCACCAUCUCA 1879 UGAGAUGGUGACAAGGGUUGU [1325-1345](21/21) 1380 GUACAACCCUUGUCACCAUCU 1880 AGAUGGUGACAAGGGUUGUAC [1323-1343](21/21) 1381 GACACACUACUUGGCCUCAGU 1881 ACUGAGGCCAAGUAGUGUGUC [1215-1235](21/21) 1382 CUGAUCUAUCACUUUGCAAAG 1882 CUUUGCAAAGUGAUAGAUCAG [1072-1092](21/21) 1383 CCUGCUACUUUAAGCCAUUCA 1883 UGAAUGGCUUAAAGUAGCAGG [1017-1037](21/21) 1384 CGAGAAGUAUUUGACUUCAGU 1884 ACUGAAGUCAAAUACUUCUCG [213-233](21/21) 1385 UUCCCGUUUGUAGAUGACAAU 1885 AUUGUCAUCUACAAACGGGAA [492-512](21/21) 1386 GCGCAGACAUUCCCGUUUGUA 1886 UACAAACGGGAAUGUCUGCGC [483-503](21/21) 1387 UGGAUCUGCCAACUACUCCCA 1887 UGGGAGUAGUUGGCAGAUCCA [404-424](21/21) 1388 GCUCAGUUACAACUAGAUGAA 1888 UUCAUCUAGUUGUAACUGAGC [327-347](21/21) 1389 GCAAAAUCAUAGCCAAAACUA 1889 UAGUUUUGGCUAUGAUUUUGC [2033-2053](21/21) 1390 CUGAGCUAGUUUUUUUGUACU 1890 AGUACAAAAAAACUAGCUCAG [1954-1974](21/21) 1391 GGAGGAUUUGACCUUUUCUGA 1891 UCAGAAAAGGUCAAAUCCUCC [1937-1957](21/21) 1392 UAGGAGGAUUUGACCUUUUCU 1892 AGAAAAGGUCAAAUCCUCCUA [1935-1955](21/21) 1393 GCUGCUCAGAAUUGCAGAAAA 1893 UUUUCUGCAAUUCUGAGCAGC [1641-1661](21/21) 1394 GUGGCUGCUCAGAAUUGCAGA 1894 UCUGCAAUUCUGAGCAGCCAC [1638-1658](21/21) 1395 AAGUGGCUGCUCAGAAUUGCA 1895 UGCAAUUCUGAGCAGCCACUU [1636-1656](21/21) 1396 UGGUCCUAAAACACCAGUACA 1896 UGUACUGGUGUUUUAGGACCA [1286-1306](21/21) 1397 GAAUGGUCCUAAAACACCAGU 1897 ACUGGUGUUUUAGGACCAUUC [1283-1303](21/21) 1398 GGCCUCAGUGAUUCUGAAGUG 1898 CACUUCAGAAUCACUGAGGCC [1227-1247](21/21) 1399 UCUUCCAGCUAUGGAGACACA 1899 UGUGUCUCCAUAGCUGGAAGA [1200-1220](21/21) 1400 GGCAUUUCACUAAACACAAGU 1900 ACUUGUGUUUAGUGAAAUGCC [1146-1166](21/21) 1401 GACUCCGGCAUUUCACUAAAC 1901 GUUUAGUGAAAUGCCGGAGUC [1140-1160](21/21) 1402 UGCUACUUUAAGCCAUUCACU 1902 AGUGAAUGGCUUAAAGUAGCA [1019-1039](21/21) 1403 GUAUUUGACUUCAGUCAGCGA 1903 UCGCUGACUGAAGUCAAAUAC [219-239](21/21) 1404 UGAGCCCAGUAUCAGCAACAG 1904 CUGUUGCUGAUACUGGGCUCA [986-1006](21/21) 1405 GAGAAGUAUUUGACUUCAGUC 1905 GACUGAAGUCAAAUACUUCUC [214-234](21/21) 1406 UCGAGAAGUAUUUGACUUCAG 1906 CUGAAGUCAAAUACUUCUCGA [212-232](21/21) 1407 CAGUGAACUCAUUAAAUUCAG 1907 CUGAAUUUAAUGAGUUCACUG [913-933](21/21) 1408 AGAAGCCAAACUGACAGAAGU 1908 ACUUCUGUCAGUUUGGCUUCU [761-781](21/21) 1409 GGGAGGAGCUAUUAUCCAUUC 1909 GAAUGGAUAAUAGCUCCUCCC [676-696](21/21) 1410 UUGGGAGGAGCUAUUAUCCAU 1910 AUGGAUAAUAGCUCCUCCCAA [674-694](21/21) 1411 GCCCCUGUUGAUUUAGACGGU 1911 ACCGUCUAAAUCAACAGGGGC [630-650](21/21) 1412 GACAGGUGAAUUUCUCCCAAU 1912 AUUGGGAGAAAUUCACCUGUC [350-370](21/21) 1413 UGCAAAAUCAUAGCCAAAACU 1913 AGUUUUGGCUAUGAUUUUGCA [2032-2052](21/21) 1414 UCCCAAAAGUAAGAAGCCAGA 1914 UCUGGCUUCUUACUUUUGGGA [1895-1915](21/21) 1415 UUGUUCCCAAAAGUAAGAAGC 1915 GCUUCUUACUUUUGGGAACAA [1891-1911](21/21) 1416 ACAACUCCAAAAGGAGCAAGA 1916 UCUUGCUCCUUUUGGAGUUGU [293-313](21/21) 1417 UCCCAUUCCCUGUAGAAAAAA 1917 UUUUUUCUACAGGGAAUGGGA [1510-1530](21/21) 1418 UCAUCGGAAAACCCCAUUCAC 1918 GUGAAUGGGGUUUUCCGAUGA [1418-1438](21/21) 1419 CAGAGAAAGAAUUGCCUGUAA 1919 UUACAGGCAAUUCUUUCUCUG [1390-1410](21/21) 1420 CAGAGCACUCACGUGCAUGAU 1920 AUCAUGCACGUGAGUGCUCUG [1350-1370](21/21) 1421 CACCAGUACAUUCUUCUGGGG 1921 CCCCAGAAGAAUGUACUGGUG [1297-1317](21/21) 1422 UCCGGCAUUUCACUAAACACA 1922 UGUGUUUAGUGAAAUGCCGGA [1143-1163](21/21) 1423 UGACUCCGGCAUUUCACUAAA 1923 UUUAGUGAAAUGCCGGAGUCA [1139-1159](21/21) 1424 CCCUCACCUGCUACUUUAAGC 1924 GCUUAAAGUAGCAGGUGAGGG [1011-1031](21/21) 1425 GUCCAGAAGCCAAACUGACAG 1925 CUGUCAGUUUGGCUUCUGGAC [757-777](21/21) 1426 GCUGGUUGAGACUACCAUGGU 1926 ACCAUGGUAGUCUCAACCAGC [731-751](21/21) 1427 UUCCUGAGUUACAGUGUCUUA 1927 UAAGACACUGUAACUCAGGAA [694-714](21/21) 1428 CCCCUGUUGAUUUAGACCGUA 1928 UACCGUCUAAAUCAACAGGGG [631-651](21/21) 1429 CCAGUGGAUCUGCCAACUACU 1929 AGUAGUUGGCAGAUCCACUGG [400-420](21/21) 1430 AGACAGGUGAAUUUCUCCCAA 1930 UUGGGAGAAAUUCACCUGUCU [349-369](21/21) 1431 UCGCUCAGUUACAACUAGAUG 1931 CAUCUAGUUGUAACUGAGCGA [325-345](21/21) 1432 CCAGAUGUUAAGAAAAACUAG 1932 CUAGUUUUUCUUAACAUCUGG [1911-1931](21/21) 1433 AGCCAGAUGUUAAGAAAAACU 1933 AGUUUUUCUUAACAUCUGGCU [1909-1929](21/21) 1434 AUGGCAAUGUUUUCCUUGUUC 1934 GAACAAGGAAAACAUUGCCAU [1876-1896](21/21) 1435 AGAGAUGGCAAUGUUUUCCUU 1935 AAGGAAAACAUUGCCAUCUCU [1872-1892](21/21) 1436 GAACAACUCCAAAAGGAGCAA 1936 UUGCUCCUUUUGGAGUUGUUC [291-311](21/21) 1437 GGGCAAAAGCUCUCCAUAUCC 1937 GGAUAUGGAGAGCUUUUGCCC [1492-1512](21/21) 1438 AUCGGAAAACCCCAUUCACAA 1938 UUGUGAAUGGGGUUUUCCGAU [1420-1440](21/21) 1439 ACGUGCAUGAUGCCCAAUGUG 1939 CACAUUGGGCAUCAUGCACGU [1360-1380](21/21) 1440 GGUCCUAAAACACCAGUACAU 1940 AUGUACUGGUGUUUUAGGACC [1287-1307](21/21) 1441 CAGUCAGCGACGGAAAGAGUA 1941 UACUCUUUCCGUCGCUGACUG [230-250](21/21) 1442 CUCCGGCAUUUCACUAAACAC 1942 GUGUUUAGUGAAAUGCCGGAG [1142-1162](21/21) 1443 GACUUCAGUCAGCGACGGAAA 1943 UUUCCGUCGCUGACUGAAGUC [225-245](21/21) 1444 AAGCCAUUCACUCUCUGAACU 1944 AGUUCAGAGAGUGAAUGGCUU [1028-1048](21/21) 1445 CUGAGCCCAGUAUCAGCAACA 1945 UGUUGCUGAUACUGGGCUCAG [985-1005](21/21) 1446 GACAGUGAACUCAUUAAAUUC 1946 GAAUUUAAUGAGUUCACUGUC [911-931](21/21) 1447 GAAGACCCCAACCAGUUGACA 1947 UGUCAACUGGUUGGGGUCUUC [894-914](21/21) 1448 AGCCAAACUGACAGAAGUUGA 1948 UCAACUUCUGUCAGUUUGGCU [764-784](21/21) 1449 AAGCUGGUUGAGACUACCAUG 1949 CAUGGUAGUCUCAACCAGCUU [729-749](21/21) 1450 GAGGAGCUAUUAUCCAUUCCU 1950 AGGAAUGGAUAAUAGCUCCUC [678-698](21/21) 1451 GGUCCACAGCUCAUCAUGAUG 1951 CAUCAUGAUGAGCUGUGGACC [99-119](21/21) 1452 GCUACUAAUCAGGCUCAGUCA 1952 UGACUGAGCCUGAUUAGUAGC [585-605](21/21) 1453 CCCAGUCUUCAUUGCUACUAA 1953 UUAGUAGCAAUGAAGACUGGG [572-592](21/21) 1454 UGAUGACUGCAUGCAGCUUUU 1954 AAAAGCUGCAUGCAGUCAUCA [461-481](21/21) 1455 GAGACAGGUGAAUUUCUCCCA 1955 UGGGAGAAAUUCACCUGUCUC [348-368](21/21) 1456 UUUCGCUCAGUUACAACUAGA 1956 UCUAGUUGUAACUGAGCGAAA [323-343](21/21) 1457 AUGAUAUGACAUCUGGCUAAA 1957 UUUAGCCAGAUGUCAUAUCAU [2238-2258](21/21) 1458 GGACACCAUUUGGGCUAGUUU 1958 AAACUAGCCCAAAUGGUGUCC [2142-2162](21/21) 1459 UCAUAGCCAAAACUAGUAUAG 1959 CUAUACUAGUUUUGGCUAUGA [2039-2059](21/21) 1460 UCUGAGCUAGUUUUUUUGUAC 1960 GUACAAAAAAACUAGCUCAGA [1953-1973](21/21) 1461 GAGAUGGCAAUGUUUUCCUUG 1961 CAAGGAAAACAUUGCCAUCUC [1873-1893](21/21) 1462 GUGAAUACUCCCUGCAGCAAA 1962 UUUGCUGCAGGGAGUAUUCAC [1849-1869](21/21) 1463 AACCUUAUUCUCCUAGUGAAU 1963 AUUCACUAGGAGAAUAAGGUU [1834-1854](21/21) 1464 GGAAAACCUUAUUCUCCUAGU 1964 ACUAGGAGAAUAAGGUUUUCC [1830-1850](21/21) 1465 GCCUUCACCUACUGAAAAAAC 1965 GUUUUUUCAGUAGGUGAAGGC [1759-1779](21/21) 1466 UGCUCAGAAUUGCAGAAAAAG 1966 CUUUUUCUGCAAUUCUGAGCA [1643-1663](21/21) 1467 GAGUAUGAGCUGGAAAAACAG 1967 CUGUUUUUCCAGCUCAUACUC [246-266](21/21) 1468 AUGGUCCUAAAACACCAGUAC 1968 GUACUGGUGUUUUAGGACCAU [1285-1305](21/21) 1469 ACACACUACUUGGCCUCAGUG 1969 CACUGAGGCCAAGUAGUGUGU [1216-1236](21/21) 1470 GAGACACACUACUUGGCCUCA 1970 UGAGGCCAAGUAGUGUGUCUC [1213-1233](21/21) 1471 AUGGAGACACACUACUUGGCC 1971 GGCCAAGUAGUGUGUCUCCAU [1210-1230](21/21) 1472 AGCUAUGGAGACACACUACUU 1972 AAGUAGUGUGUCUCCAUAGCU [1206-1226](21/21) 1473 GCAUUUCACUAAACACAAGUC 1973 GACUUGUGUUUAGUGAAAUGC [1147-1167](21/21) 1474 UCAGUCAGCGACGGAAAGAGU 1974 ACUCUUUCCGUCGCUGACUGA [229-249](21/21) 1475 AUUCACUCUCUGAACUUCUAA 1975 UUAGAAGUUCAGAGAGUGAAU [1033-1053](21/21) 1476 AACACAGAUUUUGGUGAUGAA 1976 UUCAUCACCAAAAUCUGUGUU [945-965](21/21) 1477 GUCAACACAGAUUUUGGUGAU 1977 AUCACCAAAAUCUGUGUUGAC [942-962](21/21) 1478 GAACUCAUUAAAUUCAGAUGC 1978 GCAUCUGAAUUUAAUGAGUUC [917-937](21/21) 1479 ACCAGUUGACAGUGAACUCAU 1979 AUGAGUUCACUGUCAACUGGU [904-924](21/21) 1480 AGGUAACUGUAGUCCACAUUU 1980 AAAUGUGGACUACAGUUACCU [830-850](21/21) 1481 UAUACCCUCAAUGGAAAAAGA 1981 UCUUUUUCCAUUGAGGGUAUA [806-826](21/21) 1482 UCCAUUCCUGAGUUACAGUGU 1982 ACACUGUAACUCAGGAAUGGA [690-710](21/21) 1483 GGAGCUAUUAUCCAUUCCUGA 1983 UCAGGAAUGGAUAAUAGCUCC [680-700](21/21) 1484 ACGGUAUGCAACAGGACAUUG 1984 CAAUGUCCUGUUGCAUACCGU [646-666](21/21) 1485 CCCUGUUGAUUUAGACGGUAU 1985 AUACCGUCUAAAUCAACAGGG [632-652](21/21) 1486 GUAGCCCCUGUUGAUUUAGAC 1986 GUCUAAAUCAACAGGGGCUAC [627-647](21/21) 1487 GAAACUUCUGUUGCUCAGGUA 1987 UACCUGAGCAACAGAAGUUUC [609-629](21/21) 1488 GAGAGCCCAGUCUUCAUUGCU 1988 AGCAAUGAAGACUGGGCUCUC [567-587](21/21) 1489 AAACCAGUGGAUCUGCCAACU 1989 AGUUGGCAGAUCCACUGGUUU [397-417](21/21) 1490 UUAGGAGGAUUUGACCUUUUC 1990 GAAAAGGUCAAAUCCUCCUAA [1934-1954](21/21) 1491 AAGCCAGAUGUUAAGAAAAAC 1991 GUUUUUCUUAACAUCUGGCUU [1908-1928](21/21) 1492 UUCCCAAAAGUAAGAAGCCAG 1992 CUGGCUUCUUACUUUUGGGAA [1894-1914](21/21) 1493 GGAGCAAGAGAAAGCCUUUUU 1993 AAAAAGGCUUUCUCUUGCUCC [305-325](21/21) 1494 GAUGGCAAUGUUUUCCUUGUU 1994 AACAAGGAAAACAUUGCCAUC [1875-1895](21/21) 1495 AAGGAGCAAGAGAAAGCCUUU 1995 AAAGGCUUUCUCUUGCUCCUU [303-323](21/21) 1496 AAGCCUUCACCUACUGAAAAA 1996 UUUUUCAGUAGGUGAAGGCUU [1757-1777](21/21) 1497 UGAAGCUCAACUUGCAUUAAU 1997 AUUAAUGCAAGUUGAGCUUCA [1586-1606](21/21) 1498 AUGAAGCUCAACUUGCAUUAA 1998 UUAAUGCAAGUUGAGCUUCAU [1585-1605](21/21) 1499 CCCAUUCCCUGUAGAAAAAAU 1999 AUUUUUUCUACAGGGAAUGGG [1511-1531](21/21) 1500 UUGGAGGCUCAUCUCACAAGA 2000 UCUUGUGAGAUGAGCCUCCAA [1461-1481](21/21) 1501 AGCUGGAAAAACAGAAAAAAC 2001 GUUUUUUCUGUUUUUCCAGCU [253-273](21/21) 1502 GUGCAUGAUGCCCAAUGUGAG 2002 CUCACAUUGGGCAUCAUGCAC [1362-1382](21/21) 1503 CACGUGCAUGAUGCCCAAUGU 2003 ACAUUGGGCAUCAUGCACGUG [1359-1379](21/21) 1504 ACACCAGUACAUUCUUCUGGG 2004 CCCAGAAGAAUGUACUGGUGU [1296-1316](21/21) 1505 GUCCUAAAACACCAGUACAUU 2005 AAUGUACUGGUGUUUUAGGAC [1288-1308](21/21) 1506 UGGAGACACACUACUUGGCCU 2006 AGGCCAAGUAGUGUGUCUCCA [1211-1231](21/21)

Example 2 In Vitro Testing of siRNA Compounds for Nrf2

1. General

1.5-2×10⁵ tested cells (HeLa or 293 cells) were seeded per well in a 6-well plate (70-80% confluent).

24 h subsequently, cells were transfected with siRNA oligos using lipofectamine 2000 reagent (Invitrogene) at a final concentration of 500 μM, 5 nM, 20 nM or 40 W. The cells were incubated at 37° C. in a CO2 incubator for 72 h.

As positive control for cell transfection, PTEN-Cy3 labeled siRNA oligos were used.

As negative control for siRNA activity, GFP siRNA oligos were used.

72 h after transfection cells were harvested and RNA was extracted from cells.

Transfection efficiency was tested by fluorescent microscopy.

Results:

The percent of inhibition of gene expression using specific preferred siRNAs was determined using qPCR analysis of target gene in cells expressing the endogenous gene. The data in Table C demonstrate the percent of knockdown of the expression of the target gene in cells. In general, the siRNAs having specific sequences that were selected for in vitro testing were specific for both human and the rat/rabbit genes. Similar results of reduced expression of specific genes are obtained with other siRNAs, the sequences of which are listed in Tables A and B.

TABLE C Percent of knockdown of the expression of the target Nrf2 human gene in cells using siRNAs having preferred sequences (results of separate experiments are indicated). % knockdown of Active siRNA Sequence the human gene NFE2L2_10 Sense: 20 nM in HeLa cells: (Sense SEQ ID NO: 501) CCCUGUCGAAAAAAUCAUU 91%, 54%, 51% having one mismatch (in bold) Antisense: inhibition compared to the human AAUGAUUUUUUCGACAGGG 5 nM in HeLa cells: sequence (Antisense  91% inhibition SEQ ID NO: 1004) NFE2L2_4 Sense: 20 nM - 80%, 41% (Sense SEQ ID NO: 119) CCCAUUCACAAAAGACAAA inhibition (Antisense  Antisense: 5 nM - 73%, 36% SEQ ID NO: 622) UUUGUCUUUUGUGAAUGGG inhibition NFE2L2_5 Sense: 20 nM - 69%, 17% (Sense SEQ ID NO: 195): CAGCAGGACAUGGAUUUGA inhibition (Antisense  Antisense: 5 nM - 47%, 16% SEQ ID NO: 698) UCAAAUCCAUGUCCUGCUG inhibition NFE2L2_9 Sense: 20 nM - 44% (Sense SEQ ID NO: 502) UCCCUGUCGAAAAAAUCAU inhibition (Antsense Antisense: 5 nM - 25% SEQ ID NO: 1005) AUGAUUUUUUCGACAGGGA inhibition NFE2L2_11 Sense: 20 nM - 46% (Sense SEQ ID NO: 503) CCUGUCGAAAAAAUCAUUA inhibition (Antisense Antisense: 5 nM - 23% SEQ ID NO: 1006) UAAUGAUUUUUUCGACAGG inhibition NFE2L2_1 Sense: 20 nM - 55%, 73%, (Sense SEQ ID NO: 2) GGAGGGGUAAGAAUAAAGU 53% inhibition (Antisense Antisense: SEQ ID NO: 505) ACUUUAUUCUUACCCCUCC NFE2L2_2 Sense: 20 nM - 92%, 49%, (Sense SEQ ID NO: 7) GCCCUCACCUGCUACUUUA 46% inhibition (Antisense Antisense: SEQ ID NO: 510) UAAAGUAGCAGGUGAGGGC NFE2L2_3 Sense: 20 nM - 96%, 83%, (Sense SEQ ID NO: 10) UCCCGUUUGUAGAUGACAA 82% inhibition (Antisense Antisense: SEQ ID NO: 513) UUGUCAUCUACAAACGGGA NFE2L2_12 Sense: 20 nM - 95%, 96%, (Sense SEQ ID NO: 26) GUAAGAAGCCAGAUGUUAA 76% inhibition (Antisense Antisense: SEQ ID NO: 529) UUAACAUCUGGCUUCUUAC

Example 3 The Effect of Nrf2 siRNA Treatment on Tumor Growth In Vivo

Methods:

Tumor Xenografts: A549 cells (5×10⁶) were injected into the hind leg of male athymic nude mice and the tumor was measured weekly. The tumor volumes were measured using the following formula: [length (mm)×width (mm)×width (mm)×0.52]. In the lung metastasis experiments, 2×106 A549-C8-luc cells were injected into SCID-Beige mice (Charles River, Mass.) intravenously.

For the in vivo experiments, all siRNA compounds were chemically synthesized being stabilized by alternating 2-O′-Me modifications. The sequence of siRNA targeting human Nrf2 used for in vivo experiments is 5′-UCCCGUUUGUAGAUGACAA-3′ (sense, SEQ ID NO:10) and 5′-UUGUCAUCUACAAACGGGA-3′ (antisense, SEQ ID NO:513). The sequence of control siRNA targeting GFP is 5′-GGCUACGUCCAGGAGCGCACC-3′ (sense, SEQ ID NO: 2007) and 5′-GGUGCGCUCCUGGACGUAGCC-3′ (antisense, SEQ ID NO: 2008).

For in vivo delivery of siRNA into subcutaneous tumors, siRNA duplexes diluted in PBS were injected into the hind leg tumors using insulin syringes at a concentration of 10 μg/mm³. Intraperitoneal injections of carboplatin were given at a dose of 40 mg/kg body weight. Both siRNA and carboplatin were administered twice weekly for 4 weeks.

For lung tumor delivery, female C57B6 mice were injected with Lewis Lung Carcinoma (LLC) cells (0.5×10⁶) intravenously, 24 days prior to the delivery experiment. Upon development of lung metastases, mice were administered with 100 μg/mouse of Cy3-labeled naked chemically stabilized reference siRNA via nebulizer inhalation on 3 consequent days. Mice were euthanized 24 hrs after the last inhalation. Upon termination, lungs were inflated with ice-cold 4% paraformaldehyde, followed by manual sectioning with razor blades. Clearly visible large surface tumors were sectioned separately. Resulting sections were analyzed by Bio-Rad Confocal microscope using a 20× Water objective and 2× zoom combined to give a total of 40× magnification. Control, non-siRNA-treated lungs were used to set up background fluorescence level.

For aerosol delivery of Nrf2 or GFP siRNA into lung tumors, 100 μg of siRNA duplex diluted in PBS was aerosolized using a nebulizer. Mice were given three dose of siRNA (100 μg/dose) every week, for 4 weeks, using a nebulizer. Intraperitoneal injections of carboplatin were given at a dose of 30 mg/kg body weight twice/week.

In Vivo Imaging: Animals inoculated with A549-C8-luc cells, which express a luciferase reporter gene, were anesthetized and injected intraperitoneally with 250 ul of luminescent substrate (15 mg/ml stock) D-Luciferin Firefly (Xenogen Cat# XR-1001). The animals were then imaged and analyzed by using the Xenogen IVIS Optical Imaging Device in the Johns Hopkins Oncology Center.

Statistical Analysis—Statistical comparisons were performed by Student's t-tests. A value of p<0.05 was considered statistically significant. Tumor weights and changes in tumor volume were summarized using descriptive statistics. Differences in tumor measures between treatment groups were examined using linear regression models with generalized estimating equations (GEE). The distributions of both tumor measurements were skewed, so log transformations were used.

Results:

A tumor xenograft experiment with A549 cells which were injected into the hind leg was conducted in order to test the anti-tumor activity of Nrf2 siRNA in vivo. Mice bearing subcutaneous tumors were treated with Nrf2 siRNA (comprising SEQ ID NO: 10) by direct injection into the tumor and by carboplatin twice a week for 4 weeks and tumor weight was measured at the termination of the experiment. Treatment with control non-targeting siRNA (GFP siRNA) did not inhibit tumor growth as compared to control mice treated with PBS alone. The tumor weights were significantly different between GFP siRNA and Nrf2 siRNA treated tumors (P=0.0002). Treatment with Nrf2 siRNA alone reduced mean tumor weight by 53% (±20% SD) compared to the control group. When Nrf2 siRNA was combined with carboplatin, there was an even greater reduction in mean tumor weight in all animals. To explore the effect of radiation exposure in combination with Nrf2 inhibition in vivo, the same A549 cell tumor xenograft model was used. In comparison with control siRNA+ionizing radiation (IR) treated tumors, combination of Nrf2 siRNA+IR (2 fractions of 3Gy irradiation) produced an additive effect on tumor growth inhibition (Tables D1 and D2). Delivery of naked Nrf2 siRNA duplex into tumor inhibited the expression of Nrf2 and its downstream target genes (HO-1 and GCLm), P<0.05 (FIG. 1).

TABLE D1 Mean (SD) of tumor weights and changes in tumor volume in tumor xenograft mice following treatment with Nrf2 siRNA (SEQ ID NO: 10), carboplatin, radiation and combination thereof (Experiment 1). Mean (SD) change in Mean (SD) tumor Treatment tumor volume (mm³) weight (mg) GFP siRNA 532.94 (260.94)  426 (187.83) GFP siRNA + 246.77 (189.39) 341.25 (275.27)   radiation GFP siRNA + 352.34 (170.96) 212 (66.48) carboplatin Nrf2 siRNA 249.17 (111.37) 238 (64.58) Nrf2 siRNA + 116.04 (147.9)  118 (70.85) radiation Nrf2 siRNA + 58.78 (80.06) 110 (57.62) carboplatin

TABLE D2 Mean (SD) of tumor weights and changes in tumor volume in tumor xenograft mice following treatment with Nrf2 siRNA (SEQ ID NO: 10), carboplatin, radiation and combination thereof (Experiment 2). Mean (SD) change in Mean (SD) tumor Treatment tumor volume (mm³) weight (mg) GFP siRNA   375 (233.7)  327.5 (201.56) GFP siRNA + 187.33 (76.66)    185 (67.16) carboplatin Nrf2 siRNA 138.67 (128.24) 143.33 (90.74)  Nrf2 siRNA +   45 (33.81) 58.33 (21.37) carboplatin

The delivery of naked siRNA duplexes into orthotopic lung tumors was examined. Mice were injected with Lewis lung carcinoma cells and 24 days later (when the mice developed larger tumors) mice were inhaled for three consecutive days with 100 μg/day/mouse of Cy3 labeled naked chemically stabilized reference siRNA using a nebulizer. Twenty four hours after last siRNA administration, mice were sacrificed; lungs harvested and sectioned. Resulting sections were analyzed by Bio-Rad Confocal microscope using a 20× Water objective and 2× zoom combined to give a total of 40× magnification. Control, non-siRNA-treated lungs were used to set up background fluorescence level. The fluorescence results revealed localization of Cy3 labeled siRNA in a large surface tumor, especially in intraparenchymal tumor. The large surface-protruding tumors showed Cy3 signal but the intensity was several folds lower than that observed in the small intra-parenchymal tumors.

In the next step, mice with orthotopic growing tumors expressing ARE-Luc Nrf2-dependent reporter were used. The mice with A549 lung tumors were treated with Nrf2 siRNA (SEQ ID NO: 10) intranasally using a nebulizer followed by carboplatin treatment. As demonstrated in FIG. 2, Nrf2 siRNA administered intranasally inhibited Nrf2 reporter activity in vivo, indicating specific inhibition of Nrf2 expression using Nrf2 siRNA following intranasal administration. Mice receiving Nrf2 siRNA together with carboplatin demonstrated higher growth inhibition as compared with control mice receiving GFP siRNA together with carboplatin (Table D3). The mean lung weight at termination and the luminescent flux intensity (evaluated by in vivo imaging) were lowest in mice treated with Nrf2 siRNA+carboplatin. Thus, combination of Nrf2 siRNA with carboplatin/radiation led to a reduction in tumor growth after 4 weeks of treatment compared with either agent alone. This study suggests that Nrf2 siRNA inhibitors may be considered as highly efficient promoters for the antineoplastic potential of platinum drugs such as carboplatin, causing additive/synergistic effects in lung cancer cells.

TABLE D3 Mean (SD) of lung tumor weights following intranasal treatment with Nrf2 siRNA (SEQ ID NO: 10) alone, and in combination with carboplatin. Treatment Mean (SD) tumor weight (mg) GFP siRNA 710 (167.93) GFP siRNA + 480 (95.13) carboplatin Nrf2 siRNA 802 (172.25) Nrf2 siRNA + 370 (62.05) carboplatin Vehicle siRNA 760 (98.99) Vehicle siRNA + 495 (49.5) carboplatin 

The invention claimed is:
 1. A double-stranded RNA compound having the structure: 5′ (N)_(x)-Z 3′ (antisense strand) 3′ Z′-(N′)_(y )5′ (sense strand)

wherein each N and N′ is a ribonucleotide which may independently be modified or unmodified in its sugar residue; wherein each of (N)_(x) and (N′)_(y) is an oligomer in which each consecutive N or N′ is joined to the next N or N′ by a covalent bond; wherein each of x and y is an integer between 19 and 40; wherein each of Z and Z′ may be present or absent, but if present is 1-5 consecutive nucleotides covalently attached at the 3′ terminus of the strand in which it is present; and wherein the sequences of (N)_(x) and (N′)_(y) are complementary and the sequence of (N)_(x) comprises SEQ ID NO. 622 and the sequence of (N′)_(y) comprises SEQ ID NO:119.
 2. The compound of claim 1, wherein the covalent bond is a phosphodiester bond.
 3. The compound of claim 2, wherein x=y=19.
 4. The compound of claim 3, wherein Z and Z′ are both absent.
 5. The compound of claim 3, wherein one of Z or Z′ is present.
 6. The compound of claim 1, wherein all of the ribonucleotides in (N)_(x) or (N′)_(y) or both are unmodified in their sugar residues.
 7. The compound of claim 1, wherein at least one ribonucleotide in (N)_(x) or (N′)_(y) comprises a modification in its sugar residue.
 8. The compound of claim 7, wherein the modification comprises a modification at the 2′ position of the sugar residue.
 9. The compound of claim 8, wherein the modification at the 2′ position of the sugar residue results in the presence of a moiety selected from the group consisting of amino, fluoro, alkoxy and alkyl groups.
 10. A composition comprising a pharmaceutically acceptable carrier and a compound of claim
 1. 11. A method of treating a patient suffering from a disorder comprising administering to the patient the compound of claim 1 in a therapeutically effective dose thereby treating the patient.
 12. The method of claim 11, wherein the disorder is a cancerous disease selected from the group consisting of lung cancer, breast cancer, cervical cancer, colon cancer, gastric cancer, kidney cancer, leukemia, liver cancer, lymphoma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, sarcoma, skin cancer, testicular cancer, and uterine cancer.
 13. The method of claim 12, wherein the cancerous disease is a lung cancer.
 14. The method of claim 13, wherein the compound comprises a naked siRNA compound.
 15. The method of claim 14, wherein the compound is administered via aerosol directly to the inner lung of the patient.
 16. The method of claim 15, further comprising administering to the patient a chemotherapy drug.
 17. The compound of claim 9, wherein the modification at the 2′ position of the sugar residue comprises the presence of an alkoxy group.
 18. The compound of claim 17, wherein the alkoxy group is a methoxy (2′-O-methyl) group.
 19. The compound of claim 18, wherein alternating ribonucleotides in the antisense strand or in the sense strand or both strands are modified in their sugar residues.
 20. The compound of claim 19, wherein each ribonucleotide at the 5′ terminus and at the 3′ terminus in the antisense strand is modified in its sugar residue, and each ribonucleotide at the 5′ terminus and at the 3′ terminus of the sense strand is unmodified in its sugar residue.
 21. The compound of claim 20, wherein each of the antisense strand and the sense strand is non-phosphorylated at its 3′ terminus and at its 5′ terminus.
 22. A double-stranded RNA compound having the structure:

wherein each of A, C, G and U is an unmodified or a 2′O-methyl sugar modified ribonucleotide and each consecutive ribonucleotide is joined to the next ribonucleotide by a covalent bond; and wherein each vertical line represents base pairing between the ribonucleotides.
 23. The compound of claim 20, wherein each of the antisense strand and the sense strand is phosphorylated at its 3′ terminus.
 24. The compound of claim 23, wherein in the compound the ribonucleotide at the 5′ terminus of both the antisense strand and the sense strand is non-phosphorylated.
 25. The compound of claim 22, wherein in the compound a 2′-O-methyl group is present on the sugar reside of the first, third, fifth, seventh, ninth, eleventh, thirteenth, fifteenth, seventeenth and nineteenth ribonucleotide of the antisense strand, and wherein a 2′-O-methyl group is present on the sugar residue of the second, fourth, sixth, eight, tenth, twelfth, fourteenth, sixteenth and eighteenth ribonucleotide of the sense strand.
 26. The composition of claim 10, wherein the compound is present in an amount effective to down-regulate expression of a human nuclear factor erythroid-2 related factor 2 (Nrf2) gene. 